Fig. 5
TanIIA regulated autophagic related genes protein expression and signaling pathway in A375 cells. A375 cells were treated with TanII A (0, 1, 2, or 4 μg/mL) for 48 h in 6 well plate (5 × 105 cell/2 ml). The cells protein lysates were harvested for Western blot. The primary antibodies used for the Western blots included mouse anti-Beclin-1, mouse anti-LC3 -II, mouse anti-PI3K, mouse anti- phosphorylated (p) - protein kinase B (P-Akt), p- mammalian target of rapamycin (P-mTOR), p-p70S6K1. The GAPDH was used as internal protein level control. a. The Western blot images for anti-Beclin-1, LC3 -II, PI3K, P-Akt, P-mTOR, p-p70S6K1, and GAPDH in A375 cells treated with Tan II A. Each concentration showed two duplicate treatments. b. Tan II A increased Beclin-1 protein expression in a dose dependent manner. c. Tan II A enhanced LC3-II protein expression in a dose dependent manner. d. Tan II A decreased PI3K protein expression in a dose dependent manner. e. Tan II A reduced P-Akt protein phosphorylation in a dose dependent manner. f. Tan II A inhibited P-mTOR protein phosphorylation in a dose dependent manner. g. Tan II A decreased P-p70S6K1 protein phosphorylation in a dose dependent manner. Each bar was the average of three replicates (n = 3). *P < 0.05, ** P < 0.01, and ***P < 0.001 for comparison of indicated Tan II A treatments to the negative control (0 μg/mL Tan II A) respectively