Fig 1: Fisetin suppresses mTOR kinase activity and reduces phosphorylated mTOR pathway components in cytokine-activated human epidermal keratinocytes. (A) ELISA analysis of mTOR kinase activity in control, fisetin-treated, or cytokine [rhIL-6/IL-22 (15ng/ml each)]-treated primary keratinocytes lysates. (B) Western blot analysis of differential protein expression levels of p-AktSer473, p-AMPK (Thr172)?, p-mTORSer2448, and p-raptor (Ser792) in fisetin (15µM) treated, rapamycin (100nM) or (IL-17A+TNFa)-activated HEKa compared with untreated control cells. (C) Graphs of the semi-quantified intensity of bands for p-Akt, p-AMPK, p-mTOR, and p-raptor. Bar represents means ± SD of three separate experiments each performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 vs. control for IL-6/IL-22-treated only or vs. IL-22 or IL-17A+TNFa for fisetin-treated cells. ns, not significant.