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. 2016 Oct 14;7(50):82864–82875. doi: 10.18632/oncotarget.12655

Figure 6. Effects of plumbagin on human hepatic stellate cells.

Figure 6

A. Plumbagin promoted pAMPK levels in a time-dependent manner. LX-2 cells were treated with 10 μM plumbagin for the indicated times, and pAMPK levels were analyzed. B. Plumbagin inhibited pSTAT3, p-p65, pAkt and p-mTOR levels in a dose-dependent manner. However, plumbagin had no effect on pSMAD2. LX-2 cells were first treated with the indicated concentrations of plumbagin for 1 h after incubation. Whole cells were harvested and analyzed using Western blots for pAMPK, pSTAT3, p-p65, pAkt, p-mTOR and pSMAD2. GAPDH was used as a loading control. C. Plumbagin facilitated the bind of AMPK with p300. LX-2 cells were treated with 10 μM plumbagin for 1 h, and cell lysates were immunoprecipitated using an anti-AMPK antibody, followed by immunoblotting with an anti-p300 or anti-AMPK antibody. D. Plumbagin decreased mRNA level of fibrosis-associated genes. LX-2 cells were treated with plumbagin (200 ng/ml) or TGF-β1 (2 ng/ml) or combination for 24 h, and then total RNA were extracted for quantitative PCR of COL-1, COL-3 and α-SMA. Representative images are shown for all panels. Data are expressed as means ± SD. *p< 0.05; **p< 0.01. p values were calculated using a two-sided Student's t test.