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. 2022 Jun 16;54:102366. doi: 10.1016/j.redox.2022.102366

Fig. 3.

Fig. 3

MerTK modulates bioenergetics metabolism through promoting the Warburg Effect and suppressing OXPHOS in HCC cells. (A and C) The intact cellular ECAR of control, MerTK stable knockdown (A) and overexpression (C) MHCC97H and HCCLM3 cells were measured in real time using the Seahorse XF96 Extracellular Flux Analyzer. Data were analyzed by Seahorse XF-96 Extracellular Flux Analyzer. ECAR is reported in mpH/minute, the results were normalized to cell number. Data are presented as mean ± SD, statistical significance was assessed by One-way ANOVA with Bonferroni post-test, *P < 0.05, **P < 0.01. (B and D) The intact cellular OCR of control, MerTK stable knockdown (B) and overexpression (D) MHCC97H and HCCLM3 cells were measured in real time using the Seahorse XF96 Extracellular Flux Analyzer. Data were analyzed by Seahorse XF-96 Extracellular Flux Analyzer. OCR is reported in pmols/minute, the results were normalized to cell number. Data are presented as mean ± SD, statistical significance was assessed by One-way ANOVA with Bonferroni post-test, *P < 0.05, **P < 0.01. (E) Glucose uptake of MerTK stable knockdown or overexpression MHCC97H, HCCLM3 by 2-NBDG incorporation by FACS. Data are presented as mean ± SD, statistical significance was assessed by One-way ANOVA with Bonferroni post-test and Student's t-test, *P < 0.05, **P < 0.01. (F) Lactate production of MerTK stable knockdown or overexpression MHCC97H, HCCLM3 cells were measured following manufacturer's instructions. Data are presented as mean ± SD, statistical significance was assessed by One-way ANOVA with Bonferroni post-test and Student's t-test, *P < 0.05, **P < 0.01. (G and H) Immunoblots of glycolysis related enzymes PGK1, LDHA, PFKM, PKM2, PDHK1 in MerTK stable knockdown (G) or overexpression (H) MHCC97H and HCCLM3 cells. β-Actin was used as a loading control.