Fig. 4

The expression of PHKA2 and its effects on glycolipid metabolism in GBM. A PHKA2 protein levels were analyzed in NBTs, LGGTs, and HGGTs by western blot. Data presented as mean ± SD (n = 3, each group), ^*P < 0.05 versus NBTs group; ^**P < 0.01 versus NBTs group; ^##P < 0.01 versus LGGTs group. B PHKA2 protein levels in NHA, U251, and U373 cells were detected via western blot. Data presented as mean ± SD (n = 3, each group), ^**P < 0.01 versus the NHA group. C, D The extracellular acidification rate (ECAR) was measured to demonstrate the effects of PHKA2 on glycolysis in U251 and U373 cells, and the glycolysis was calculated. Data presented as mean ± SD (n = 3, each group), ^**P < 0.01 versus PHKA2(–)NC group; ^##P < 0.01 versus PHKA2( +)NC group. E, F Lactate production and glucose uptake were measured in U251 and U373 cells after PHKA2 knockdown or overexpression. Data presented as mean ± SD (n = 3, each group), ^**P < 0.01 versus PHKA2(–)NC group; ^##P < 0.01 versus PHKA2( +)NC group. G, H Intracellular triglyceride and cholesterol expression levels were measured to evaluate the effect of PHKA2 on lipogenesis. Data presented as mean ± SD (n = 3, each group), ^**P < 0.01 versus PHKA2(–)NC group; ^##P < 0.01 versus PHKA2( +)NC group. I The effect of PHKA2 on proliferation was analyzed via CCK-8 assay. ^**P < 0.01 versus PHKA2(–)NC group; ^##P < 0.01 versus PHKA2( +)NC group. Data presented as mean ± SD (n = 3, each group). J Representative confocal fluorescence imaging of lipid droplets (LDs) stained by BODIPY 493/503 (green) in U251 and U373 cells. The nucleus (blue) was stained by DAPI. Scale bars = 10 µm. Data presented as mean ± SD (n = 15, each group). ^**P < 0.01 versus PHKA2(–)NC group; ^##P < 0.01 versus PHKA2( +)NC group. Statistical analysis was performed using the one-way ANOVA method