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. 2020 Sep 23;7(5):ENEURO.0170-20.2020. doi: 10.1523/ENEURO.0170-20.2020

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Copyright © 2020 Nguyen et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 3. — Tppp KO mice display defective fear conditioning. A, Fear conditioning assay experimental design. On day 0, mice were acclimated to a wired-floor cage with a background scent of 10% ethanol. On day 1, the mice were again acclimated to the cage and then a cue sound was played for 20 s followed by a foot shock and a 60-s ITI. The cue, trace, shock, and ITI were repeated for a total of four times. On day 2, mice were placed in the same wired floor cage with 10% ethanol scent. On day 3, mice were acclimated to a cage with solid smooth flooring and 1% vanilla background scent and then given the cue sound, without the shock; n = 11 mice per genotype for all data in this figure. Detailed statistical results can be found in Extended Data Figure 3-1; power analyses can be found in Extended Data Figures 1-2, 2-2. B, On day 1 (training), Tppp KO mice had similar freezing responses as wild-type mice, with no significant differences in percentage time spent freezing when evaluated by two-way ANOVA with post hoc Sidak’s multiple comparisons test. Graphs represent significance based on two-way ANOVA with post hoc Sidak’s multiple comparisons test. Evaluating wild-type versus Tppp KO means as independent t tests (i.e., at different time points/periods) gives p values of 0.371 (wild type: 2.306%, Tppp KO: 0.472%) at baseline and of 0.042 (wild type: 62.94%, Tppp KO: 47.79%) during ITI4 (p = 0.042). All other p values generated by independent t tests were >0.05. C, On day 2 (context recall), Tppp KO mice displayed less freezing overall compared with wild-type mice. p = 0.007 by Welch’s t test. D, On day 3 (context recall), Tppp KO mice displayed significantly less freezing than wild-type mice at all timepoints during the first two rounds of training. Graphs represent significance based on two-way ANOVA with post hoc Sidak’s multiple comparisons test. Evaluating wild-type versus Tppp KO means as independent t tests (i.e., at different time points/periods) reveals significant p values for all other timepoints except trace 3: at baseline, p = 0.0014 (wild type: 24.74%, Tppp KO: 8.650%); for cue 3, p = 0.0213 (wild type: 35.65%, Tppp KO: 13.55%); for trace 3, p = 0.0634 (wild type: 35.64%, Tppp KO: 17.06%); for ITI 3, p = 0.0473 (wild type: 31.21%, Tppp KO: 14.60%); for cue 4, p = 0.0269 (wild type: 39.66%, Tppp KO: 17.03%); for trace 4, p = 0.0294 (wild type: 38.85%, Tppp KO: 19.64%); for ITI 4, p = 0.0320 (wild type: 35.63%, Tppp KO: 18.36%). E, On day 3, Tppp KO mice displayed significantly less freezing during baseline, cue, trace, and ITI periods than wild-type mice; p values calculated by two-way ANOVA. Error bars represent SEMs. *p < 0.05, **p < 0.01, ***p < 0.001.