Fig. 4: HIFs and METTL3 are required for hypoxia-induced PLOD2 expression.
From: METTL3 facilitates renal cell carcinoma progression by PLOD2 m6A-methylation under prolonged hypoxia
A, B Expression levels of HIF-1α, HIF-2α, METTL3, and PLOD2 were determined by RT-qPCR (A) and western blotting (B) after ACHN cells were exposed to normoxia (20% O2) or hypoxia (1% O2). **P = 0.0013, 0.0022, 0.0087, and 0.0026 (left to right), ***P = 0.0002, ****P < 0.0001. C–E Examination of the knockdown efficiency when si-HIF-1α, si-HIF-1α, or double knockdown (DKD) were transfected into ACHN cells. ****P < 0.0001. F–G Silencing of HIFs decreased METTL3 and PLOD2 expression at mRNA level (F) and protein level (G) in RCC cells under prolonged hypoxia. ****P < 0.0001, *P = 0.0454 and 0.0124 (left to right), ***P = 0.0004, **P = 0.0082. (H) Luciferase reporter assay was used to determine the potential HIF-1α binding sites in METTL3 promoter region. ****P < 0.0001. I–K Expression levels of METTL3 and PLOD2 were examined in ACHN cells by RT-qPCR (I, J) and western blotting (K) when prolonged hypoxia was combined with METTL3 silencing. J *P = 0.0336 (left), 0.0274 (right), **P = 0.0010. K, *P = 0.0323, ****P < 0.0001. Data were represented as mean ± S.D. of three independent experiments.
