Figure 6
From: Multigenerational impacts of bile exposure are mediated by TGR5 signaling pathways
Adult CA-exposure alters male germ cells. (a) Testicular mRNA accumulation of Dnmt3b normalized to β-actin mRNA levels in adult F0CT;+/+ and F0CT;−/− male founders. (b) Testicular mRNA accumulation of Dnmt3b normalized to β-actin mRNA levels in adult F0CT;+/+; F0CT;−/−; F0CA;+/+ and F0CA;−/− male founders. (c) Immunohistochemistry of Smad6 and 5meC in testis of F0CT;+/+; F0CT;−/−; F0CA;+/+ and F0CA;−/− male founders. (d) Quantification of the relative percentage of seminiferous tubules with germ cells co-stained for Smad6 and 5meC in testis of F0CT;+/+; F0CT;−/−; F0CA;+/+ and F0CA;−/− male founders. (e) mRNA expression of Dnmt3b normalized to β-actin levels in GC1spg germ cell lines transfected with siGfp or siTgr5. (f) mRNA expression of Dnmt3b normalized to β-actin levels in GC1spg germ cell lines transfected with siGfp or siTgr5 and exposed to vehicle or DCA during 24 hours. (g) DNMT3B protein accumulation normalized to ACTIN levels in GC1spg germ cell lines transfected with siGfp or siTgr5 and exposed to vehicle or DCA during 24 hours. Data are expressed as means +/− SEM. n = 8–15 per group from 3 independent experiments. In a, b, c and d: *p < 0.05 vs. control diet group. In e, f and g: *Difference from the siGfp vehicle group; #difference from the siGfp DCA group. siGfp vehicle-treated cells were fixed at 100% for each siRNA condition.
