Figure 2.

ULK1 depletion increases the therapeutic effects of sorafenib in vitro and in vivo. (A) Sorafenib induced autophagy in HepG2 cells. HepG2 cells were treated with sorafenib (5 μM) for 12 h. The data are representative TEM images of three independent assays. The arrows indicate Avd. The ratio of autophagic vacuole area to the cytoplasmic area was determined by morphometric analysis. (B) The effects of 3-MA (2 mM), sorafenib (5 μM), 3-MA (2 mM) + sorafenib (5 μM) on the LC3-II levels in HepG2 cells. (C) Autophagy inhibition increased the anti-invasion effects of sorafenib in HepG2 cells. HepG2 cells were evaluated via transwell assays with the indicated treatments (sorafenib: 5 μM; 3-MA: 2 mM) for 24 h. (D, E) ULK1 silencing increased the antiproliferation effects of sorafenib. Hep3B and primary HCC cells were transfected with the indicated siRNAs. After 24 h of transfection, the cells were treated with sorafenib for 24 h, and proliferation activity was evaluated via EdU assays. (F, G) ULK1 depletion increased the therapeutic effects of sorafenib in vitro. ULK1 [Con] and ULK1 [Cas9] HepG2 cells were treated with sorafenib (5 μM) for 24 h, and proliferation and invasion activities were evaluated via CCK-8 (F) and transwell (G) assays. (H-K) ULK1 depletion increased the therapeutic effects of sorafenib in vivo. The data represent the body weight growth (H), quantified tumor weight (I), tumor growth curves (J), and fluorescence area and photographs of representative tumors (K). For panel A-G, the data are presented as representative and/or means ± s.e.m. of three independent assays. For panel H-K, the data are the mean ± s.e.m. (n = 10 per group). For panel A, an unpaired two-sided Student’s t-test was used to compare two groups. For panels B, C, D, E, F, G, H, I, J, and K, one-way ANOVA was used to compare multiple groups. *p < 0.05, **p < 0.01, ***p < 0.001.