Fig. 7: ACA suppresses the growth of β-catenin-activated HCCs.

a Representative western blots of pmTOR, mTOR, ATG5, p62, LC3B, FTH1, and TfR1 in HepG2 and SNU398 cells after treatment with ACA (0, 5, 10, 20 μM) for 48 h (n = 3 independent experiments). b Cytotoxicity of 20 μM ACA in HepG2 and SNU398 cells co-treated with sorafenib (0, 0.5, 1, 2, 4 μM) for 48 h. The gray curve was the vehicle control DMSO group, the blue curve was ACA group. Data are presented as the mean ± SEM (n = 3 independent experiments). Statistical analysis was performed using two-tailed Student’s t test. c Tumor volumes and weights of HepG2 xenografts after treatment with vehicle (Veh, 5% DMSO + 40% PEG400 + 55% saline, gray), ACA (30 mg/Kg, i.p., once every two days, blue), sorafenib (Sor, 10 mg/Kg, i.p., once every two days, red), or combination of sorafenib and ACA (Sor+ACA, 30 mg/kg ACA + 10 mg/kg Sorafenib, i.p., once every two days, orange). Data are presented as the mean ± SEM (n = 6 independent mice per group). Statistical analysis was performed using one-way ANOVA test. d Gross images and representative IHC staining images of HepG2 xenografts. Images were representative shown out of 6 independent mice per group. e Study design. Mice were subjected to hydrodynamic tail vein injection (HTVi) of c-Met/β-catenin plasmids or AKT/β-catenin plasmids. Starting from 3 weeks post injection, mice were treated with vehicle (Veh, 5% DMSO + 40% PEG400 + 55% saline) or ACA (30 mg/Kg, i.p., once every two days). f Survival curves. Mice were euthanized when they developed a high burden of liver tumors (Gray curve, vehicle control group; Blue curve, ACA-treated group). The log-rank test was used to compare overall survival between groups. g Representative gross liver images, as well as H&E and IHC staining images. Images were representative shown out at least 7 independent mice per group. Scale bar, 200 μm. h Scheme summarizing the effect of SLC13A3 inhibition on intracellular metabolic pathways that ultimately converge on autophagic ferroptosis in β-catenin-activated HCC cells. Source data are provided as a Source Data file.