Supplementary Figure 4 from Hypoxia-Induced Creatine Uptake Reprograms Metabolism to Antagonize PARP1-Mediated Cell Death and Facilitate Tumor Progression in Hepatocellular Carcinoma

Supplementary Figure 4. Creatine-mediated SERPINE1 upregulation inhibits parthanatos and aggravates HCC progression. (A) Western blot showing the regulatory effects of hypoxia and creatine/SLC6A8 on SERPINE1 expression. (B-D) CCK-8 (B), EdU incorporation (C) and colony formation assays (D) were used to determine the role of SERPINE1 in hypoxic HCC cell proliferation and colony formation. Scale bars, 50 μm. (E) Cell invasion and migration were evaluated by transwell assay. Scale bars, 50 μm. (F, G) Cell apoptosis (F) and cell cycle (G) of hypoxic control and SERPINE1-knockdown cells were measured using flow cytometry. (H) Glucose uptake of hypoxic HCC cells were measured. (I-L) The ECAR and OCR were measured in hypoxic control and SERPINE1-knockdown cells. (M) The weight statistics of subcutaneous xenografts of HCC cells. (N) IHC staining revealed the SERPINE1 expression in representative HCC samples in the TMA. Scale bars, 100 μm. (O) Correlation between SLC6A8 and SERPINE1 levels based on the TMA data. Means ± SD are shown; *P < 0.05, **P < 0.01, ***P < 0.001, and ns indicates no significant difference.