Fig. 5
PPP1R26 promotes glycolysis in HCC cells and in vivo. A Expression of PPP1R26 was evaluated by Western blot in the HepG2 and Huh7 stably expressing shRNAs (left). Beta-actin was used as a loading control. Glucose uptake and lactate production in PPP1R26-depleted HepG2 and Huh7 cells were detected (middle & right). B GFP-PPP1R26 or GFP was transfected into HepG2 and Huh7 cells. Expression of GFP-PPP1R26 was evaluated by Western blot (left). Beta-actin was used as a loading control. Glucose uptake and lactate production in HepG2 and Huh7 were detected (middle & right). C The extracellular acidification rate (ECAR) was evaluated in HepG2-PPP1R26 shRNA (left) and Huh7-PPP1R26 shRNA (middle) cells treated with glucose, oligomycin, and 2-deoxyglucose (2-DG), respectively. HepG2 and Huh7 cells were transfected with GFP or GFP-PPP1R26 (right). D The histogram analysis of glycolysis rate (ECAR after glucose injection) and glycolysis capacity (ECAR after oligomycin injection) in (C). E Huh7-PPP1R26 shRNA cells were injected subcutaneously into the left (rad arrow) and Huh7-Ctrl shRNA cells in the right (white arrow) flanks of BALB/C nude mice. PET/CT imaging was performed. Representative PET photographs of animals were shown (left). Glucose uptake in the tumor was evaluated by the average SUVmax (right). F Western blot analysis of indicated protein levels when GFP-PPP1R26 was expressed in the PKM2-depleted HCC cells (left). Glucose uptake and lactate production were detected when GFP-PPP1R26 was expressed in the PKM2-depleted HCC cells (middle & right). G Expression of PPP1R26 and GFP-PTBP1 were evaluated by Western blot in the indicated cells (left). Beta-actin was used as a loading control. ECAR was evaluated in PPP1R26-depleted Huh7 cells after GFP-PTBP1 was expressed (middle). The histogram analysis of ECAR results (right). H Expression of GFP-PPP1R26 and PTBP1 were detected by Western blot in the indicated cells (left). Beta-actin was used as a loading control. ECAR was evaluated in PTBP1-depleted Huh7 cells when GFP-PPP1R26 was expressed (middle). The histogram analysis of ECAR results (right). Data information: In (A-H), data are presented as mean ± SD. Statistical significance was assessed using two-tailed t-tests (B, D & F) or one-way ANOVA with post hoc analysis LSD test (A, D, G & H). *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001