Fig. 3
From: Placental miR-340 mediates vulnerability to activity based anorexia in mice
MiR-340 exerts its effects through targeting of GR, Cry2, and H3f3b. a−f Validation of potential miR-340 target in prenatal stress (PNS) vs. control female placentas showed that Sirt7 and Ythdf3 were not significantly correlated with miR-340 (a, f). b−e In contrast, GR, Hdac9, Cry2, and H3f3b expression was significantly higher in PNS and inversely correlated with miR-340. g−i Placental GR, CRY2, and H3F3b protein levels were increased by PNS (N = 4). j Lentiviral KD of miR-340 in BeWo choriocarcinoma cells reduced the expression of miR-340 (F(1,7) = 5.654, p = 0.050, N = 4). k MiR-340 KD did not significantly affect mRNA levels of the target genes. l At the protein level, miR-340 KD increased GR (t(6) = 3.687, p = 0.010) and CRY2 (t(6) = 3.156, p = 0.0197), and tended to increase H3F3b. m Within the control group, miR-340 expression shows great endogenous variability (N = 28, from six dams). n Female fetuses surrounded by two males in the uterus show lower levels of placental miR-340 compared to females surrounded by females or mixed sexes (one-way ANOVA F(2,23) = 9.31, p = 0.0013). Data are presented in mean and s.e.m. and Tukey’s multiple comparison tests. o−q The “high” and “low” subgroups included samples above 1.3 and below 0.70 relative to the groups’ average respectively. The expression of GR, Cry2, and H3f3b inversely correlated with the endogenous levels of miR-340. Data are presented as mean and s.e.m. r Fetal hypothalamic gene expression of AN candidate genes differed between the “high” and “low” subgroups (F(6,5) = 13.1, p = 0.006). Data are presented in mean and s.e.m. (N = 6)
