Fig 2: The effect of LZG on lipid accumulation, cell injury, and PPARα-regulated gene expression in AML12 cells induced by fatty acid. (a) The lipid accumulation indicated by Nile red staining. (b) The cell viability. (c) The mRNA expression of EHHADH relative to β-actin. (d) Protein expression of ACOX1 relative to β-actin. n = 3; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.

Fig 3: AMPK-PROX1 axis impairs tumourigenesis and dictates therapeutic response.a–c Analysis of Prox1f/f (WT) and Alb-Cre; Prox1f/f (Prox1-cKO) mice with DEN-induced liver cancer with normal or high BCAA (200%) diets. Representative images of livers (a) and the number of tumours (b), and the Liver/body weight (c) of the mice (n = 5) as indicated. Scale bar, 1 cm. d–f Representative images and H&E staining of livers (d) and the number of tumours (e), and the Liver/body weight (f) of the mice (n = 5) as indicated. g Huh7 cells stably expressing the indicated siRNAs were subcutaneously injected into in nude mice respectively with or without metformin (500 mg/L) treatment. Shown are average tumour volumes over time (n = 7). Data are presented as mean ± SEM. h SMCC-7721 cells stably overexpressing the PROX1 variants were subcutaneously injected into nude mice respectively with or without metformin (500 mg/L) treatment as indicated, and the weights of tumours are shown (n = 6). i Schematic model of lung tumourigenesis from KrasG12D/Lkb1L/L mice treated with a lenti-Cre-PROX1 virus. Eight weeks after nasal inhalation, mice were killed and analysed. j–k Representative H&E image (j) of lung tumour are shown. Scale bar, 500 µm. The tumour burden (k) and average tumour numbers (l) were calculated and plotted (n = 5 mice for each group). m Immunohistochemistry analysis of the levels of Prox1, p-S6K, Bckdhb, Acadsb and Ehhadh in the above tissues as indicated. Scale bar, 50 µm. n–p Representative the H&E image (n) of lung tumour from KrasG12D/Lkb1L/L mice treated with a lenti-Cre-sgPROX1 and sgSCR virus as indicated with or without phenformin (1.5 g/L) treatment (n = 5 mice for each group). Scale bar, 500 µm. Statistical analysis of the tumour burden (o) and average tumour numbers (p) were shown. q, r Representative IHC staining images (q) and statistical data (r) of Ki67 expression in the above tissues (n = 5) as indicated. Scale bar, 50 µm. For b–c, e–f, h, k–l, o–p and r, data represent the mean ± SD. Statistical significance was assessed using two-tailed unpaired Student’s t-test. n.s. not significant. Source data are provided as a Source Data file.