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. Author manuscript; available in PMC: 2010 Aug 23.
Published in final edited form as: Cancer Res. 2007 Mar 15;67(6):2497–2507. doi: 10.1158/0008-5472.CAN-06-3075

Figure 4. Leptin fails to stimulate proliferation of hepatocellular carcinoma cells in the presence of inhibitors of JAK/STAT, ERK, or AKT.

Figure 4

A to C, HepG2 and Huh7 cells were cultured, serum-starved for 16 h, and treated with leptin (100 ng/mL; L). For combined treatment, cells were pretreated with 100 µmol/L AG490 (A or A + L), 10 µmol/L PD98059 (P or P + L), or 10 µmol/L LY294002 (Ly or Ly + L) for 45 min followed by leptin treatment. Untreated controls (U). Cell lysates were prepared and quantified for protein content. A total of 100 µg protein was resolved on 10% SDS-PAGE followed by immunoblot analysis with specific antibodies against total or phosphorylated forms of STAT3 (A), ERK (B), and AKT (C). The representative histogram is the densitometric analysis of bands showing fold increase in levels of phosphorylated forms with respect to total protein. *, P < 0.01, compared with untreated control cells; #, P < 0.01, compared with leptin treatment; **, P < 0.05, leptin with inhibitor compared with respective inhibitor treatment alone. D, cells were treated as described earlier, and BrdUrd incorporation analysis was done as described in Materials and Methods. Columns, mean of experiments done in triplicates thrice; bars, SE. *, P < 0.001, compared with untreated control cells; #, P < 0.005, compared with leptin treatment. Immunoblot for β-actin was done as a loading control.