Additional file 2: of Phloretin attenuates STAT-3 activity and overcomes sorafenib resistance targeting SHP-1–mediated inhibition of STAT3 and Akt/VEGFR2 pathway in hepatocellular carcinoma

Figure S2. PH potentiates the effect of Sor in vitro in HCCs. (A) HepG2, SK-Hep1 and Hep 3B2. 1-7 cells were treated with PH or Sor or both at indicated doses for 48 h and growth inhibition was assessed by MTT assay. (B) PH enhanced DNA fragmentation. PH or Sor or both at indicated doses for 48 h and DNA fragmentation was measured by cell death detection ELISA. (C) Apoptosis was analyzed by annexin V labeling and FACS analysis. Annexin V (+) cells were quantified. (D) pSTAT-3 activity was measured by ELISA. (E) The protein levels of p-STAT3 was determined by western blot (F) HCC cells were exposed PH or Sor or both at the indicated doses for 48 h and SHP- 1 phosphatase activity was determined. (G) Purified dN1 and D61A mutants of SHP-1 were insensitive to PH treatment. (H) Percentage of apoptotic cells was analyzed by flow cytometry in purified dN1 and D61A mutants of SHP-1 treated with PH. Experiments were conducted in triplicate and mean values ± SD (bars) are shown. *p<0.05, and **p<0.01 versus control. (DOCX 161 kb)