Figure 4
E2/ESR1 signaling inhibits hepatic lipogenesis. Representative images of liver sections from Control (A) and LERKO (B) mice after staining with Oil Red O as a measure of lipid accumulation (magnification: ×20). n = 3 per group. (C) Quantification of Oil Red O staining using Image (J). (D) Hepatic triglyceride levels in Control and LERKO mice. (E,F) mRNA levels of hepatic lipogenic genes Fas and Acc1 in Control and LERKO mice were measured by q-RT-PCR. (C–F) *p < 0.05 versus Control, **p < 0.01 versus Control. Data from Fig. 4E,F are representative of results obtained from 6–10 mice in each group. (G,H). Primary hepatocytes from Control and LERKO mice were treated with vehicle or different doses of E2 (10−10 M, 10−9 M, 10−8 M) or GPER agonist G-1 (10−10 M, 10−9 M, 10−8 M), mRNA levels of Fas and Acc1 were measured by q-RT-PCR. *p < 0.05 versus vehicle, **p < 0.01 versus vehicle, ***p < 0.001 versus vehicle. (I,J) Primary hepatocytes from Control and LERKO mice were treated with vehicle or different doses of E2 (10−10 M, 10−9 M, 10−8 M) in absence or in presence of GPER antagonist G-15 (10−8 M), mRNA levels of Fas and Acc1 were measured by q-RT-PCR. *p < 0.05 versus vehicle, **p < 0.01 versus vehicle, ***p < 0.001 versus vehicle. (K,L) ChIP assay experiments were performed with liver tissues using antibody to ESR1, or with rabbit preimmune serum (IgG) and primers flanking the Fas (K) and Acc1 (L) promoters. Real-time PCR data with an inset of a 1.5% agarose gel as a representative example. Results were normalized to input and shown as fold enrichment IgG from 3 independent ChIP experiments. The experiments were performed 2 weeks after virus injection. Values are means ± SD. *p < 0.05 versus IgG, ***p < 0.001 versus IgG.
