Fig. 4. CREB knockout (KO) in PFC increases susceptibility but is rescued by Zfp189 overexpression.

a) Experimental timeline to evaluate behavioral effects of CREB KO in PFC. b) Local KO of CREB in PFC produces social avoidance in SI in a subthreshold social defeat procedure (mixed model ANOVA, interaction F_1,26 = 4.656, p = 0.040, n=11,17 mice, Bonferroni post-test p < 0.001). c-d) CREB KO reduces mRNA levels of (c) Creb1 (two-tailed t test, t = 2.974, p = 0.006) and (d) Zfp189 (two-tailed Mann-Whitney, U = 45.0, p = 0.036), n=11,16 mice. e) Experimental timeline to determine whether Zfp189 overexpression in PFC reverses the deleterious effects of CREB KO. f) CREB KO in PFC produces social avoidance in the SI test but concurrent overexpression of Zfp189 reverses this deficit (mixed model ANOVA, target F_1,25 = 5.690, p = 0.025, n=13,14 mice, Bonferroni post-test p < 0.05). g) Overexpression of Zfp189 in PFC increases sucrose preference in CREB KO mice (two-tailed t test, t = 5.176, p < 0.001, n=13 mice). h) Experimental schematic for female sub-chronic variable stress (SCVS) to investigate behavioral effects of CREB KO and Zfp189 overexpression in PFC. i) CREB KO in PFC increases latency to eat in the novel arena, but not when CREB KO is paired with Zfp189 overexpression (mixed model ANOVA, interaction F_1,35 = 5.301, p = 0.027, n=9,11,10,10 mice, Bonferroni post-test compared to CREB⁻Zfp189^- in novel arena, p < 0.01 for CREB⁺Zfp189^-, p < 0.01 for CREB⁻Zfp189⁺, and p < 0.05 for CREB⁺Zfp18⁺). j) Mice with local KO of CREB in PFC have a lower sucrose preference than control mice, an effect blocked by concurrent Zfp189 overexpression (Kruskall-Wallis test, χ²(3) = 8.475, p = 0.037, n=9,11,10,10 mice, two-tailed Mann-Whitney post-test p = 0.009 for local KO of CREB and p = 0.029 for overexpression of Zfp189 with concurrent CREB KO). *p < 0.05, **p < 0.01, ***p < 0.001. Bar graphs show mean ± SEM.