Figure 5. Inducible Silencing of Klf9 Expression in Forebrain Excitatory Neurons of Female Mice Prevents CRS-Induced Potentiation of Contextual Fear Acquisition.

(A) Schematic representation of tTS:Tre-Klf9 female mice with different doxycycline diet schedules and chronic restraint stress timeline.

(B) CFCDL procedure in which mice were trained to discriminate between a footshock delivered in context A and safe context B.

(C) Freezing behavior on day 1. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 0.13, NS; doxycycline F_(1,40) = 0.97, NS; interaction F_(1,40) = 0.04, NS.

(D) Freezing behavior on day 2. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 0.49, NS; doxycycline F_(1,40) = 0.28, NS; interaction F_(1,40) = 3.64, NS.

(E) Freezing behavior on day 3. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 1.62, NS; doxycycline F_(1,40) = 9.35, p < 0.01; interaction F_(1,40) = 5.4, p < 0.05; *p < 0.05 and **p < 0.01, ON CORT ON DOX versus OFF DOX.

(F) Freezing behavior on day 4A. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 0.01, NS; doxycycline F_(1,40) = 6.29, p < 0.05; interaction F_(1,40) = 0.27, NS.

(G) Freezing behavior on day 4B. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 0.46, NS; doxycycline F_(1,40) = 1.14, NS; interaction F_(1,40) = 2.56, NS.

(H) Discrimination ratio calculated for contexts A and B. Data (mean ± SEM; n = 12, 11, 10, and 11 mice per group) were analyzed using mixed-factor two-way ANOVA: CRS F_(1,40) = 1.16, NS; doxycycline F_(1,40) = 0.03, NS; interaction F_(1,40) = 1.99, NS.

See also Figures S1 and S2.