Table 2. Summary of statistical analyses applied in this study.
| Figure | Dataset | Groups | n-number* | Statistical test | p-value |
|---|---|---|---|---|---|
| 1C | Cumulative number of DCV fusion events per neuron plotted against time | control DKO |
3 (36) 3 (34) |
Two-way repeated measures ANOVA |
ptime x genotype = 0.4823 ptime <0.0001 (****) pgenotype = 0.3166 |
| 1D | Total number of DCV fusion events/neuron | control DKO |
3 (36) 3 (34) |
Mann-Whitney test | p=0.0970 |
| 1E | Number of intracellular NPY-pHluorin puncta/neuron | control DKO |
3 (36) 3 (35) |
Unpaired t test | p=0.0224 (*) |
| 1F | Fraction of released DCVs (% total pool) | control DKO |
3 (36) 3 (34) |
Two-way repeated measures ANOVA |
ptime x genotype >0.9999 ptime <0.0001 (****) pgenotype = 0.9014 |
| 1G | Fluorescence intensity peaks (averaged per neuron) | control DKO |
3 (36) 3 (34) |
Mann-Whitney test | p=0.4509 |
| 1H | Band intensities of syntaxin-1 complexes (WB) | DKO (% control) |
4 cultures | One sample t-test (compare to 100%) |
p37kDa = 0.2858 p80 kDa = 0.5714 p200 kDa = 0.0002 (***) |
| 1 suppl 1E | Dendrite length/ neuron | control DKO |
3 (30) 3 (28) |
Unpaired t-test | p=0.5902 |
| 1 suppl 1 F | Number of synaptophysin puncta/ mm dendrite | control DKO |
3 (30) 3 (28) |
Unpaired t-test | p=0.1380 |
| 1 suppl 1 G | Intensity of synaptophysin signal in neurites | control DKO |
3 (30) 3 (28) |
Unpaired t-test | p=0.6561 |
| 1 suppl 3 A | Band intensities of SNAP25 complexes (WB) | DKO (% control) | 3 cultures | One sample t-test (compare to 100%) |
p25kDa = 0.7265 p80 kDa = 0.8454 p200 kDa = 0.0064 (**) |
| 1 suppl 3B | Band intensities of VAMP2 complexes (WB) | DKO (% control) | 3 cultures | One sample t-test (compare to 100%) |
p15kDa = 0.0585 p80 kDa = 0.4283 |
| 2B | Intensity of NPY-pHluorin (ICC) | control DKO rescue |
3 (30) 3 (29) 3 (29) |
One-way ANOVA | p<0.0001 (****) |
| Tukey’s multiple comparisons post hoc test |
pcontrol vs DKO <0.0001 (****) pDKO vs rescue <0.0001 (****) |
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| Intensity of tomosyn (ICC) | control DKO rescue |
3 (30) 3 (30) 3 (30) |
One-way ANOVA | p<0.0001 (****) | |
| Tukey’s multiple comparisons post hoc test |
pcontrol vs DKO <0.0001 (****) pDKO vs rescue <0.0001 (****) |
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| 2D | Intensity of BDNF (ICC) | control DKO rescue |
3 (20) 3 (20) 3 (20) |
Kruskal-Wallis test | p=0.0009 (***) |
| Dunn’s multiple comparisons test |
pcontrol vs DKO = 0.0009 (***) pDKO vs rescue <0.0253 (*) |
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| Intensity of tomosyn (ICC) | control DKO rescue |
3 (20) 3 (20) 3 (20) |
One-way ANOVA | p<0.0001 (****) | |
| Tukey’s multiple comparisons post hoc test |
pcontrol vs DKO = 0.0012 (**) pDKO vs rescue <0.0001 (****) |
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| 2F | Band intensities of BDNF (WB) | DKO (% control) | 4 (8) | One sample t-test (compare to 100%) | p<0.0001 (****) |
| 2H | Band intensities of BDNF (WB rescue) | DKO DKO+FL DKO+ΔSNARE |
2 (4) | One sample t-test (compare each condition to 100%) |
pDKO = 0.0005 (***) pDKO+FL = 0.1224 pDKO+ΔSNARE = 0.0767 |
| 2I | Band intensities of BDNF (WB leupeptin) | control ±leupeptin DKO ±leupeptin |
2 (4) | Two-way repeated measures ANOVA |
ptreatment = 0.0640 pgenotype = 0.0240 (*) |
| 2 suppl 1B | Intensity of EGFP expressed under control of synapsin promoter | control DKO |
3 (30) 3 (28) |
Unpaired t-test | p=0.0898 |
| 2 suppl 2B | Intensity of IA-2 (neurites) | control DKO |
3 (24) 3 (24) |
Unpaired t-test | p<0.0001 (****) |
| Intensity of IA-2 (soma) | control DKO |
3 (24) 3 (24) |
Unpaired t-test | p=0.0472 (*) | |
| 2 suppl 2 C | Band intensities of IA-2 (WB) | DKO (% control) |
3 (9) | One sample t-test (compare to 100%) | p<0.0001 (****) |
| 2 suppl 3B | Intensity of LAMP1 | control DKO |
2 (10) | Unpaired t-test | p=0.9847 |
| 2 suppl 3D | Band intensities of LAMP1 (WB) | DKO (% control) |
3 (9) | One sample t-test (compare to 100%) | p=0.3716 |
| Band intensities of LIMP2 (WB) | DKO (% control) |
3 (9) | One sample t-test (compare to 100%) | p=0.7225 | |
| 4B | DCV diameter | control DKO |
3 (135 DCVs) 3 (64 DCVs) |
Unpaired t-test on means from three independent cultures | p=0.0129 (*) |
| 4E | SV diameter | control DKO |
3 (621 SVs) 3 (617 SVs) |
Unpaired t-test on means from three independent cultures | p=0.2234 |
| 4F | Number of SV/ synapse | control DKO |
3 (149 synapses) 3 (147 synapses) |
Unpaired t-test on means from three independent cultures | p=0.4713 |
| 4G | Length of active zone | control DKO |
3 (151 synapses) 3 (148 synapses) |
Unpaired t-test on means from three independent cultures | p=0.4366 |
| 4H | Length of PSD | control DKO |
3 (152 synapses) 3 (147 synapses) |
Unpaired t-test on means from three independent cultures | p=0.3752 |
| 5B | TGN area | control DKO |
3 (40) 3 (39) |
Unpaired t-test | p=0.0001 (***) |
| Nucleus area | control DKO |
3 (40) 3 (39) |
Unpaired t-test | p=0.1847 | |
| 5F | NPY-EGFP intensity in the Golgi plotted against time | control DKO |
3 (33) 3 (30) |
Two-way repeated measures ANOVA |
ptime x genotype = 0.0014 (**) ptime <0.0001 (****) pgenotype = 0.3280 |
| 5G | Time required to reach maximum NPY-EGFP intensity in the Golgi | control DKO |
3 (34) 3 (28) |
Unpaired t-test | p=0.0444 (*) |
| 5H | Rate constants of NPY-EGFP export from the Golgi | control DKO |
3 (34) 3 (30) |
Unpaired t-test | p=0.0276 (*) |
| 5J | % anterograde DCVs | control DKO |
2 (12) 2 (12) |
Unpaired t-test | p=0.9488 |
| % retrograde DCVs | control DKO |
2 (12) 2 (12) |
Unpaired t-test | p=0.7938 | |
| 5K | Speed of anterogradely trafficking DCVs | control DKO |
2 (11) 2 (12) |
Unpaired t-test | p=0.8107 |
| 6A | mRNA expression of Bdnf transcript isoforms | DKO (logFC relative to control) | 8 cultures | One sample t-test (compare to log2FC = 0) |
ptotal = 0.0003 (***) ptr1=0.0006 (***) ptr2=0.0032 (**) ptr4 <0.0001 (****) ptr6=0.1183 ptr9=0.0012 (**) |
| 6B | mRNA expression of general DCV cargos | DKO (logFC relative to control) | 8 cultures | One sample t-test (compare to log2FC = 0) |
p VgfVgf 0.0006 (***) p Pcsk1Pcsk10.0006 (***) p PamPam 0.0032 (**) p PtprnPtprn 0.0187 (*) p Scg2Scg20.0177 (*) |
| 6C | mRNA expression of general DCV cargos | DKO (logFC relative to control) | 8 cultures | One sample t-test (compare to log2FC = 0) |
p NpyNpy 0.0002 (***) p CckCck 0.0016 (**) p GrpGrp 0.0003 (***) p SstSst 0.6986 p Tac1Tac10.0201 (*) p PnocPnoc 0.0023 (**) p VipVip 0.7972 p PdynPdyn 0.8905 p PenkPenk 0.4408 p CortCort 0.0267 (*) |
| 6D | mRNA expression of DCV cargos upon tomosyn re-expression | DKO DKO +tom |
3 cultures | Paired t-test on log2FC |
p Bdnf-tr1Bdnf-tr10.5453 p Bdnf-tr4Bdnf-tr40.4470 pVgf = 0.6284 p Pcsk1Pcsk10.4625 p GrpGrp 0.2974 |
| 6E | Effect of Cre on DCV cargo mRNA expression in Stxbp5/5llox and WT neurons | ΔCre - Stxbp5/5llox Cre - Stxbp5/5llox |
3 cultures | Paired t-test on log2FC |
p Bdnf-tr1Bdnf-tr10.0274 (*) pVgf = 0.0115 (*) p Pcsk1Pcsk10.0039 (**) p GrpGrp 0.0062 (**) p GapdhGapdh 0.2759 |
| ΔCre - WT Cre - WT |
3 cultures | Paired t-test on log2FC |
p Bdnf-tr1Bdnf-tr10.0160 (*) pVgf = 0.0057 (**) p Pcsk1Pcsk10.0040 (**) p GrpGrp 0.0041 (**) p GapdhGapdh 0.3046 |
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| 6G | Band intensities of BDNF (WB) | ΔCre Cre |
3 (3) | Paired t-test | p=0.0476 (*) |
| Band intensities of tomosyn (WB) | ΔCre Cre |
3 (3) | Paired t-test | p=0.7470 | |
| 6 suppl 2B | Intensity of NPY-pHluorin (ICC) | ΔCre Cre |
3 (18) | Unpaired t-test | p=0.8022 |
| Intensity of tomosyn (ICC) | ΔCre Cre |
3 (18) | Unpaired t-test | p=0.7105 |
*
n-number is the number of independent culture preparations. The number of individual observations (e.g. neurons or lanes) is provided in brackets.