Figure 5
From: The Role of Bone Morphogenetic Protein Signaling in Non-Alcoholic Fatty Liver Disease
BMP-induced DGAT2 (diacylglycerol O-acyltransferase 2) expression is mediated through SMAD signaling. HepG2 cells treated with siRNA directed against SMAD 1/5 had reduced SMAD 1/5 protein levels (A) and decreased canonical BMP signaling as evidenced by decreased mRNA expression of ID1 (inhibitor of DNA binding 1) when stimulated with the BMP ligand BMP2 (B). BMP2-induced DGAT2 expression is SMAD1/5 dependent (C). Using chromatin immunoprecipitation combined with qPCR (ChIP-qPCR), BMP2 stimulation resulted in increased binding of SMAD 1/5 to a putative SMAD promoter region (1000 base pairs upstream of the DGAT2 start site) at 10-fold higher levels compared to control levels (D). Experiments had five replicates per group for panels (B,C) and three replicates per group for panel (D). SMAD western blots were run on 2 separate gels given similar protein weights. Comparisons were performed using 1-way ANOVA with Sidak’s multiple comparison testing.
