Fig 2: Membrane-bound and total level ratio of HRas and RhoA protein content in brain areas of Fmr1-Δexon 8 and WT adult (PND90) male rats. The figure illustrates a typical Western blot and a graph representing the ratio of the densitometric analysis between membrane-bound and total levels of HRas (top) and RhoA (bottom) in PND90 rats. The loading control was performed by using caveolin 1 (Figure S3). (A): amygdala; (B): prefrontal cortex (cortex); (C): nucleus accumbens. Values represent mean ± SD obtained from 6 animals performed in duplicate. Statistical analysis was carried out by using the unpaired Student’s t-test. ** = p < 0.01 vs. WT.

Fig 3: Impact of age and CR on membrane-bound and total levels of HRas and RhoA in rat cortex. Representative Western blots and densitometric analysis of membrane-bound (m-) and total content (t-) of HRas (a) and RhoA (b) in rat cortex. Vinculin and caveolin served as housekeeping proteins to normalize protein loading. 3M (3-month-old rats); 24M (24-month-old rats); ND (normal diet); CR (caloric intake reduced by 40% for the last 6 months of life). Statistical analysis was performed by using one-way ANOVA followed by Tukey’s post hoc test. * = p < 0.05, *** = p < 0.001, vs. 3M ND.

Fig 4: RhoA and Ras activation in the brain of rat fetuses in utero exposed to 2.5 µg/kg/day of BPA. Representative Western blots of membrane-bound RhoA (a) and Ras (b) and their content in total lysate of fetuses brain from prenatally BPA exposed and CTR animals. The graphs represent the ratio of membrane-bound and total levels of the proteins in brain lysates. n = 6 for each experimental group. Statistical analysis was carried out by using two-way ANOVA followed by Bonferroni post-hoc test. ** = p < 0.01 vs. CTR M; *** = p < 0.001 vs. CTR M; # = p < 0.05 vs. CTR F; ## = p < 0.01 vs. CTR F.