Figure 1, behavior characterization |
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Social approach |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.0001 |
F(3,118) = 7.344 |
Preference |
p < 0.0001 |
F(1,118) = 52.56 |
Genotype |
p = 0.2661 |
F(3,118) = 1.336 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Novel animal, novel object |
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Control VEH n = 18 |
p = 0.0022 |
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Mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 17 |
p < 0.0001 |
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Mutant RAPA n = 14 |
p < 0.0001 |
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Social novelty |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.2465 |
F(3,112) = 1.400 |
Preference |
p < 0.0001 |
F(1,112) = 23.17 |
Genotype |
p = 0.1095 |
F(3,112) = 2.060 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Novel animal, novel object |
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Control VEH n = 17 |
p = 0.0199 |
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Mutant VEH n = 12 |
p > 0.9999 |
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Control RAPA n = 17 |
p = 0.0059 |
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Mutant RAPA n = 14 |
p = 0.0051 |
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Social olfaction |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p < 0.0001 |
F(42,735) = 2.641 |
Scent |
p < 0.0001 |
F(14,735) = 39.46 |
Group |
p < 0.0001 |
F(3,735) = 37.52 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Water |
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Control VEH n = 19 vs mutant VEH n = 12 |
p = 0.8170 |
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Control RAPA n = 12 vs mutant RAPA n = 10 |
p > 0.9999 |
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Control VEH n = 19 vs control RAPA n = 12 |
p > 0.9999 |
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Mutant VEH n = 12 vs mutant RAPA n = 10 |
p > 0.9999 |
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Almond |
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Control VEH n = 19 vs mutant VEH n = 12 |
p > 0.9999 |
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Control RAPA n = 12 vs mutant RAPA n = 10 |
p > 0.9999 |
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Control VEH n = 19 vs control RAPA n = 12 |
p = 0.3913 |
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Mutant VEH n = 12 vs mutant RAPA n = 10 |
p = 0.1125 |
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Social scent |
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Control VEH n = 19 vs mutant VEH n = 12 |
p = 0.0149 |
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Control RAPA n = 12 vs mutant RAPA n = 10 |
p > 0.9999 |
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Control VEH n = 19 vs control RAPA n = 12 |
p = 0.0010 |
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Mutant VEH n = 12 vs mutant RAPA n = 10 |
p < 0.0001 |
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Grooming |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.7009 |
F(1,55) = 0.1491 |
Treatment |
p = 0.6617 |
F(1,55) = 0.1935 |
Group |
p < 0.0001 |
F(1,55) = 19.07 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Control VEH n = 10 vs mutant VEH n = 14 |
p = 0.0067 |
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Control RAPA n = 18 vs mutant RAPA n = 17 |
p = 0.0053 |
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Control VEH n = 10 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 17 |
p > 0.9999 |
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Water Y maze |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p < 0.0001 |
F(15,378) = 3.671 |
Day |
p < 0.0001 |
F(5,378) = 83.08 |
Group |
p < 0.0001 |
F(3,378) = 12.10 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Trial 1 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p > 0.9999 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Trial 2 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p > 0.9999 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Trial 3 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p > 0.9999 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Rev. trial 1 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p < 0.0001 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p < 0.0001 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Rev. trial 2 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p = 0.0186 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p = 0.0846 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Rev. trial 3 |
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Control VEH n = 20 vs mutant VEH n = 17 |
p > 0.9999 |
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Control RAPA n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 17 vs mutant RAPA n = 13 |
p > 0.9999 |
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Rotarod |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p < 0.0001 |
F(12,390) = 3.767 |
Day |
p < 0.0001 |
F(4,390) = 16.40 |
Group |
p < 0.0001 |
F(3,390) = 70.61 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Day 1 |
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Control VEH n = 25 vs mutant VEH n = 15 |
p = 0.1112 |
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Control RAPA n = 19 vs mutant RAPA n = 23 |
p = 0.9056 |
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Control VEH n = 25 vs control RAPA n = 19 |
p > 0.9999 |
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Mutant VEH n = 15 vs mutant RAPA n = 23 |
p = 0.0071 |
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Day 2 |
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Control VEH n = 25 vs mutant VEH n = 15 |
p = 0.0001 |
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Control RAPA n = 19 vs mutant RAPA n = 23 |
p = 0.4920 |
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Control VEH n = 25 vs control RAPA n = 19 |
p > 0.9999 |
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Mutant VEH n = 15 vs mutant RAPA n = 23 |
p < 0.0001 |
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Day 3 |
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Control VEH n = 25 vs mutant VEH n = 15 |
p < 0.0001 |
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Control RAPA n = 19 vs mutant RAPA n = 23 |
p > 0.9999 |
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Control VEH n = 25 vs control RAPA n = 19 |
p = 0.1609 |
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Mutant VEH n = 15 vs mutant RAPA n = 23 |
p < 0.0001 |
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Day 4 |
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Control VEH n = 25 vs mutant VEH n = 15 |
p < 0.0001 |
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Control RAPA n = 19 vs mutant RAPA n = 23 |
p > 0.9999 |
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Control VEH n = 25 vs control RAPA n = 19 |
p = 0.0024 |
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Mutant VEH n = 15 vs mutant RAPA n = 23 |
p < 0.0001 |
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Day 5 |
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Control VEH n = 25 vs mutant VEH n = 15 |
p < 0.0001 |
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Control RAPA n = 19 vs mutant RAPA n = 23 |
p > 0.9999 |
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Control VEH n = 25 vs control RAPA n = 19 |
p < 0.0001 |
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Mutant VEH n = 15 vs mutant RAPA n = 23 |
p < 0.0001 |
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Open field |
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Two-way ANOVA |
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Distance moved |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.9653 |
F(42,989) = 0.6378 |
Minute |
p = 0.0038 |
F(14,989) = 2.318 |
Group |
p < 0.0001 |
F(3,989) = 10.53 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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0:00–1:00 (60 seconds) |
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Control VEH n = 20 vs mutant VEH n = 14 |
p = 0.6273 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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1:00–2:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p = 0.2082 |
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2:00–3:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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3:00–4:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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4:00–5:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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5:00–6:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p = 0.0411 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p = 0.8482 |
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6:00–7:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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7:00–8:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p = 0.4288 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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8:00–9:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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9:00–10:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p = 0.4031 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p = 0.0953 |
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10:00–11:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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11:00–12:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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12:00–13:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p > 0.9999 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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13:00–14:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p = 0.4511 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p = 0.7478 |
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14:00–15:00 |
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Control VEH n = 20 vs mutant VEH n = 14 |
p > 0.9999 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p > 0.9999 |
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Control VEH n = 20 vs control RAPA n = 18 |
p = 0.1904 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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Time in left |
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ANOVA table |
p value |
F
(DFn,DFd)
|
Interaction |
p = 0.9524 |
F(1,71) = 0.003591 |
Treatment |
p = 0.3550 |
F(1,71) = 0.8669 |
Genotype |
p = 0.0429 |
F(1,71) = 4.250 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Control VEH n = 20 vs mutant VEH n = 14 |
p = 0.3036 |
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Control RAPA n = 18 vs mutant RAPA n = 18 |
p = 0.2924 |
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Control VEH n = 20 vs control RAPA n = 18 |
p = 0.9581 |
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Mutant VEH n = 14 vs mutant RAPA n = 18 |
p > 0.9999 |
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Elevated plus maze |
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Two-way ANOVA |
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Open arms |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.8544 |
F(1,58) = 0.03400 |
Treatment |
p = 0.5133 |
F(1,58) = 0.4326 |
Genotype |
p = 0.0375 |
F(1,58) = 4.535 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Control VEH n = 13 vs mutant VEH n = 11 |
p = 0.4375 |
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Control RAPA n = 17 vs mutant RAPA n = 21 |
p = 0.1367 |
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Control VEH n = 13 vs control RAPA n = 17 |
p > 0.9999 |
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Mutant VEH n = 11 vs mutant RAPA n = 21 |
p > 0.9999 |
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Figure 2, cell survival |
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Cell count |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p < 0.0001 |
F(1,11) = 38.68 |
Treatment |
p < 0.0001 |
F(1,11) = 53.47 |
Genotype |
p < 0.0001 |
F(1,11) = 103.0 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Control VEH n = 4 vs mutant VEH n = 3 |
p < 0.0001 |
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Control RAPA n = 4 vs mutant RAPA n = 3 |
p = 0.0547 |
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Control VEH n = 4 vs control RAPA n = 4 |
p = 0.8911 |
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Mutant VEH n = 3 vs mutant RAPA n = 3 |
p < 0.0001 |
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Mutant VEH n = 3 vs control RAPA n = 4 |
p < 0.0001 |
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Figure 3, physiology |
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PC intrinsic firing rate |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
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Interaction |
p = 0.0225 |
F(1,142) = 5.320 |
Treatment |
p = 0.0026 |
F(1,142) = 9.406 |
Genotype |
p = 0.0001 |
F(1,142) = 15.54 |
Bonferroni's multiple comparisons test |
Adjusted p value |
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Control VEH n = 45 vs mutant VEH n = 36 |
p < 0.0001 |
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Control RAPA n = 30 vs mutant RAPA n = 35 |
p = 0.5465 |
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Control VEH n = 45 vs control RAPA n = 30 |
p > 0.9999 |
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Mutant VEH n = 36 vs mutant RAPA n = 35 |
p = 0.0005 |
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Mutant VEH n = 36 vs control RAPA n = 30 |
p < 0.0001
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PC evoked responses |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
|
Interaction |
p < 0.0001 |
F(87,1852) = 2.112 |
Current step |
p < 0.0001 |
F(29,1852) = 100.9 |
Group |
p < 0.0001 |
F(3,1852) = 313.6 |
Bonferroni's multiple comparisons test |
Adjusted p value |
|
Control VEH n = 13 vs mutant VEH n = 10 |
p < 0.0001 |
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Control RAPA n = 29 vs mutant RAPA n = 14 |
p < 0.0001 |
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Control VEH n = 13 vs control RAPA n = 29 |
p = 0.0255 |
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Mutant VEH n = 10 vs mutant RAPA n = 14 |
p = 0.0075 |
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Mutant VEH n = 10 vs control RAPA n = 29 |
p < 0.0001 |
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Membrane input resistance |
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Two-way ANOVA |
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ANOVA table |
p value |
F
(DFn,DFd)
|
Interaction |
p = 0.8539 |
F(1,86) = 0.03412 |
Treatment |
p = 1.127 |
F(1,86) = 1.254 |
Genotype |
p < 0.0001 |
F(1,86) = 18.67 |
Bonferroni's multiple comparisons test |
Adjusted p value |
|
Control VEH n = 12 vs mutant VEH n = 17 |
p = 0.0283 |
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Control RAPA n = 34 vs mutant RAPA n = 27 |
p = 0.0003 |
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Control VEH n = 12 vs control RAPA n = 34 |
p = 0.7549 |
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Mutant VEH n = 17 vs mutant RAPA n = 27 |
p = 0.9845 |
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Mutant VEH n = 17 vs control RAPA n = 34 |
p = 0.0004 |
|
Membrane capacitance |
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Two-way ANOVA |
|
|
ANOVA table |
p value |
F
(DFn,DFd)
|
Interaction |
p = 0.9078 |
F(1,23) = 0.01373 |
Treatment |
p = 0.9270 |
F(1,23) = 0.008569 |
Genotype |
p = 0.7735 |
F(1,23) = 0.08483 |
Membrane time constant |
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|
Two-way ANOVA |
|
|
ANOVA table |
p value |
F
(DFn,DFd)
|
Interaction |
p = 0.2842 |
F(1,23) = 1.203 |
Treatment |
p = 0.5662 |
F(1,23) = 0.3388 |
Genotype |
p = 0.0022 |
F(1,23) = 11.86 |
Bonferroni's multiple comparisons test |
Adjusted p value |
|
Control VEH n = 6 vs mutant VEH n = 6 |
p = 0.0114 |
|
Control RAPA n = 8 vs mutant RAPA n = 7 |
p = 0.1840 |
|
Control VEH n = 6 vs control RAPA n = 8 |
p > 0.9999 |
|
Mutant VEH n = 6 vs mutant RAPA n = 7 |
p = 0.5081 |
|
Mutant VEH n = 6 vs control RAPA n = 8 |
p = 0.0496 |
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Mouse age at time of testing (all experiments) |
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|
|
8-week treatment paradigm |
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Rotarod |
13 weeks |
Rotarod |
Open field |
14 weeks |
Open field |
Elevated plus maze |
14 weeks |
Elevated plus maze |
Social interaction (three-chamber) |
15 weeks |
Social interaction (three-chamber) |
Social olfaction |
15 weeks |
Social olfaction |
Grooming |
15 weeks |
Grooming |
Water Y maze |
16 weeks |
Water Y maze |
Immunohistochemistry |
13–16 weeks |
Immunohistochemistry |
Acute slice physiology |
13–16 weeks |
Acute slice physiology |