Figure 3

FTO regulates the LepRb-STAT3 signalling pathway in co-transfected HuH7 cells. HuH7 cells were co-transfecting with both LepRb and FTO vectors (empty vector as control) for 36 hours. Cells were then serum starved for 16 hours before three-hour leptin stimulation (100 ng/mL). A) Representative Western blots and quantitative analysis of FTO expression in co-transfected HuH7 cells, validating the FTO overexpression. B) Representative Western blots and quantitative analysis of pSTAT3(Y705) and total STAT3 in co-transfected HuH7 cells, showing that FTO represses the leptin-induced STAT3 tyrosine phosphorylation. C) Representative Western blots of pSTAT3(S727) and total STAT3 in co-transfected cells, showing that FTO represses the leptin-mediated reduction of serine phosphorylation of STAT3. D) Representative Western blots and quantitative analysis of pPKB(S473) and PKB proteins in co-tranfected HuH7 cells, showing that FTO represses the leptin-induced PKB phosphorylation. Data are means ± SEM (n = 3-6/group). *p < 0.05 compared to untreated control cells. #p < 0.05 compared to treated control cells.