Fig. 7. Hypoxia induces an ECSC phenotype via demethylation of SOX2 mRNA.

a, b Total m⁶A levels in ECSC^isk after knockdown of HIFs or ALKBH5 were determined with an m⁶A RNA Methylation Quantification Kit. c RNA protein immunoprecipitation assays detected the level of m⁶A SOX2 mRNA in ECSC^isk under normal or hypoxic conditions for 48 h treated with CoCl₂ (100 µM) or KC7F2 (100 µM), respectively. d SOX2 mRNA levels in ECSC^isk treated with CoCl₂ (100 µM) or KC7F2 (100 µM) were determined by qPCR. e RNA protein immunoprecipitation assay detected the level of m⁶A SOX2 mRNA in ALKBH5 knockdown ECSC^isk subjected to 21 or 2% O₂ for 48 h. f SOX2 mRNA levels in shALK ECSC^isk were measured by qPCR. g Determination of the level of m⁶A SOX2 mRNA in HIF knockdown ECSC^isk with or without ALKBH5 overexpression that were also subjected to 21 or 2% O₂ for 48 h. h qPCR analysis of SOX2 mRNA in Ctrl and shHIF ECSC^isk under normoxic or hypoxic conditions for 48 h. Mean ± SEM (N = 3). **^/##P < 0.01, and ***^/###P < 0.001.