Fig. 7
Ca2+ /AKT/CREB/PGC-1α axis is essential for tumor progression. A-B PGC-1α mRNA expression in the indicated cells with shCtrl or shRYR1 (A) or incubated with dantrolene or vehicle (B). C F Western blot of PGC-1α (C) proteins in AKT/CREB pathway (F) in control- or RYR1-silenced and vehicle- or dantrolene-treated ARK1 cells. D mETC genes mRNA expression in PGC-1α-silenced (siPGC-1α) or control (siCtrl) ARK1 cells transfected with RYR1 or control. E Western blot of HK1 and HK2 in control- or RYR1- silenced ARK1 cells. G Western blot of PGC-1α in ARK1-Control and ARK1-RYR1 cells treated with of AKT or CREP inhibitor vs vehicle. H Relative ATP level in control and RYR1-overexpressing cells transfected with siCtrl or siPGC-1α. *p < 0.05, **p < 0.01, ****p < 0.0001 (Student t-test). I mETC genes mRNA expression in ARK1- Control and ARK1-RYR1 cells in the presence of siPGC-1α, AKT (0.1 μM) or CREB inhibitor (0.1 nM) or controls. J Cell growth of ARK1-Control and ARK1-RYR1 incubated with AKT or CREB inhibitor. *p < 0.05, **p < 0.01, n.s., no significance (2-way ANOVA). K Western blot of phosphorylated AKT (S473), phosphorylated CREB (S133), and total AKT and CREB in ARK1-shCtrl and ARK1-shRYR1 cells treated with BAPTA or vehicle. L Graphic illustration of the molecular mechanism. In A, B, D, I, N = 3 independent experiments. HPRT was internal control. *p < 0.05, **p < 0.01, ****p < 0.0001, n.s., no significance (Student t-test). In C, E, F, G, K, N = 3 independent experiments. Data are shown as mean ± SEM. ImageJ software was applied to quantify signals. β-actin was used as loading control