Supplementary Figure 7 from Hypoxia-Induced Creatine Uptake Reprograms Metabolism to Antagonize PARP1-Mediated Cell Death and Facilitate Tumor Progression in Hepatocellular Carcinoma

Supplementary Figure 7. SERPINE1 reverses the attenuated deubiquitination activity of USP10 caused by the direct binding of PAR. (A) Western blot analysis of PKLR levels in USP10-knockdown HCC cells transfected with SERPINE1 plasmids. (B) Western blot analysis of PKLR levels in SERPINE1-knockdown HCC cells transfected with USP10 plasmids. (C, D) Co-IP assays demonstrated the effect of SERPINE1 levels on the interaction between USP10 and PKLR. (E, F) Western blot analysis of the ubiquitination of PKLR upon USP10 overexpression in HCC cells. (G) Western blotting analysis of the ubiquitination of PKLR in hypoxic HCC cells treated with MNNG or Olaparib. (H-M) The ECAR and OCR were measured in hypoxic control and PKLR knockdown cells. n = 3 independent experiments. Means ± SD are shown; *P < 0.05, **P < 0.01, ***P < 0.001, and ns indicates no significant difference.