Supplementary Figure 8 from Hypoxia-Induced Creatine Uptake Reprograms Metabolism to Antagonize PARP1-Mediated Cell Death and Facilitate Tumor Progression in Hepatocellular Carcinoma

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Supplementary Figure 8. Combining RGX-202 with lenvatinib exhibits potent antitumor effects in PDX models. (A, B) Colony formation assay was used to determine the colony formation ability of hypoxic HCC cells across different treatment groups. (C-F) Cell invasion and migration of hypoxic HCC cells across different treatment groups were evaluated by transwell assay. Scale bars, 50 μm. (G) The ECAR and OCR were measured in hypoxic HCC cells across different treatment groups. (H, I) CCK-8 (H) and EdU incorporation assays (I) were performed to evaluate the effect of SERPINE1 overexpression on cell proliferation under RGX-202 treatment. Scale bars, 50 μm. (J) Colony formation assay was used to determine the effect of SERPINE1 overexpression on colony formation ability under RGX-202 treatment. (K, L) Transwell assays were used to evaluate the effect of SERPINE1 overexpression on cell invasion and migration under RGX-202 treatment. Scale bars, 50 μm. (M, N) The ECAR and OCR were measured in hypoxic HCC cells under RGX-202 treatment with or without SERPINE1-overexpression. (O) Analysis of tumor numbers in the livers of the indicated mice (n = 5). (P) Determination of circulating ALT and AST levels in the indicated mice. (Q) Gross images of PDX model derived from low SLC6A8 expression specimens (n = 6). For cell experiments, n = 3 independent experiments. Means ± SD are shown; *P < 0.05, **P < 0.01, ***P < 0.001, and ns indicates no significant difference.

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