The stimulatory effect of OGT on lipogenic gene expression is lost when ChREBP is silenced. Mouse hepatocytes were incubated
under low glucose concentrations (G5) and adeno-infected with 5 plaque-forming units/cell of GFP, OGT, shChREBP, or OGT+shChREBP
adenovirus for 5 h. Cells were then incubated for 24 h under low glucose (G5) or high glucose concentrations plus insulin
(G25i). A: Quantitative RT-PCR analysis of OGT. Data are means ± SEM. n = 6 independent cultures. *P < 0.05 compared with GFP. B: Global O-GlcNAcylation levels and ChREBP protein content. β-Actin was used as a loading control. Representative Western blots (Wb)
are shown. n = 6 independent experiments. Lanes were run on the same gel but were noncontiguous. C: Quantitative RT-PCR analysis of L-PK, FAS, SCD1, and SREBP-1c. *P < 0.05 compared with GFP (G25i); #P < 0.05 compared with OGT (G25i). Data are means ± SEM. n = 6 independent cultures.