Fig 2: Sex- and subgroup-dependent changes in protein levels related to stress-regulating systems in the medial prefrontal cortex (mPFC) after a history of ethanol drinking and intermittent predator stress exposure. There were divergent, treatment- and subgroup-induced alterations in the proteins examined, with sex differences observed for CRF-R1, CRF-R2, and GR. Representative animals from the “sensitive” and “resilient” subgroups depicted in Figures 2, 3 were chosen for the Western blot analyses, and values were compared to separate groups of naïve mice. Values are mean ± SEM levels that were normalized to ß-actin and then normalized to the respective naïve group (dashed line) for male (A) and female (B) mice. Changes in relative protein levels were compared using normalized optical density measurements. Representative bands from the immunoblots for each protein are included beneath each panel on the graph. Representative immunoblots for each protein, brain region, and sex are depicted in Supplementary Figures 1–4. Group size: naïve (10–11 male, 8–10 female), “sensitive” subgroup (7 male, 5 female), “resilient” subgroup (10 males, 8 females). *p = 0.05, ***p = 0.001 vs. respective “resilient” subgroup; ##p = 0.01, ###p = 0.001 vs. respective naïve. The results depicted reflect either separate analyses for each sex (CRF R1, CRF R2, GR) or analyses with the sexes combined (CRF BP). See Table 2 for ANOVAs. CRF R1, corticotropin releasing factor receptor 1; CRF R2, corticotropin releasing factor receptor 2; CRF BP, corticotropin releasing factor binding protein; GR, glucocorticoid receptor.