Fig. 3. Increased DNMT3A occupancy and DNA methylation at the Fasn promoter in the late fed state are dependent on both SHP and FGF15/19.

a–c Mice were treated with vehicle or FGF19 for 2 h after fasting overnight. a Interaction of SHP with DNMTs in liver extracts determined by CoIP. b Occupancy of DNMTs at Fasn and Cyp7a1 determined by ChIP. c Occupancy of DNMT3A at the Fasn and Cyp7a1 determined by ChIP in SHP-KO or C57BL/6 mice. d FGF15-KO mice or littermates were fed normal chow for 3 h (fd) after fasting overnight (fs). Interaction of DNMT3A and SHP in liver extracts determined by CoIP (left). Input SHP in the liver determined by IB of 1% of the liver extract used for the CoIP assays (right). Tissues from 3 mice were pooled for each sample and the experiment was done twice. e–f C57BL/6 mice were fed normal chow for 3 h (fd) after fasting overnight (fs) and in (f) were fasted again overnight. e DNA methylation at specific sites of Fasn promoter determined by bisulfite sequencing. f DNA methylation at the Fasn, Cyp7a1, and Esr1 promoters determined by MeDIP. g FGF15-KO mice or littermates were fed normal chow for 3 h (fd) after fasting overnight (fs). DNA methylation at Fasn and Cyp7a1 determined by MeDIP. b, c, e, f, g Mean values ± SD are plotted (b, n = 3 mice; c, e, f, g, n = 5 mice). Statistical significance was determined by (b, c, f, g) two-way ANOVA with Tukey post-test or (e) two-tailed Student’s t test. *P < 0.05, **P < 0.01, ns, statistically not significant.