Fig. 3: circDDX21 promotes glycolysis by increasing PGAM1 expression.
From: Energy stress-induced circDDX21 promotes glycolysis and facilitates hepatocellular carcinogenesis
A Real-time RT-PCR analysis of circDDX21 and DDX21 levels in HepG2 cells transduced with lentiviruses expressing control shRNA, circDDX21 shRNA#1, or circDDX21 shRNA#2. Data shown are mean ± SD (n = 3), **p < 0.01, ns., no significance. B ECAR was measured by Seahorse XF assay in HepG2 cells expressing control shRNA, circDDX21 shRNA#1, or circDDX21 shRNA#2. The levels of glycolysis, glycolytic capacity, and glycolytic reserve were also calculated. Data shown are mean ± SD (n = 3), *p < 0.05, ***p < 0.001. C Real-time RT-PCR analysis of circDDX21 and DDX21 levels in HepG2 cells transduced with lentiviruses expressing empty vector (EV) or circDDX21. Data shown are mean ± SD (n = 3), **p < 0.01, ns., no significance. D ECAR was measured by Seahorse XF assay in HepG2 cells expressing empty vector (EV) or circDDX21. The levels of glycolysis, glycolytic capacity, and glycolytic reserve were also calculated. Data shown are mean ± SD (n = 3), *p < 0.05, **p < 0.01. E The glycolytic tracing assay was performed with 13C-labeled glucose in HepG2 cells expressing control shRNA, circDDX21 shRNA#1, or circDDX21 shRNA#2. The relative abundance of the 13C-labeled glycolytic metabolites D-glucose, pyruvate, and lactate was calculated. Data shown are mean ± SD (n = 3), *p < 0.05, ***p < 0.001. F Lysates from HepG2 cells expressing control shRNA, circDDX21 shRNA#1, or circDDX21 shRNA#2 were analyzed by western blotting to examine the protein levels of the indicated enzymes involved in the glycolysis pathway. G HepG2 cells were infected with lentiviruses expressing control, circDDX21 shRNA, or circDDX21 shRNA plus PGAM1. Forty-eight hours after infection, the ECAR was measured by a Seahorse XF assay. The levels of glycolysis, glycolytic capacity, and glycolytic reserve were also calculated. Data shown are mean ± SD (n = 3), **p < 0.01, ***p < 0.001. H HepG2 cells were infected with lentiviruses expressing control, circDDX21, PGAM1 shRNA, or both circDDX21 and PGAM1 shRNA. Forty-eight hours after infection, the ECAR was measured by a Seahorse XF assay. The levels of glycolysis, glycolytic capacity, and glycolytic reserve were also calculated. Data shown are mean ± SD (n = 3), *p < 0.05, **p < 0.01, ***p < 0.001, ns., no significance.
