Fig. 5. ALKBH5 maintains SOX2 expression by demethylating SOX2 transcripts.

a–b The SOX2 expression was detected via qRT-PCR or western blot assay after transfection with sh-ALKBH5 lentiviral vectors, *p < 0.05 compared with sh-NC group. c–d The SOX2 expression was detected via qRT-PCR or western blot assay after transfection with LV-ALKBH5 lentiviral vectors, *p < 0.05 compared with LV-NC group. e Sequence motif identified with m⁶A-seq peaks in GBM cells. f Distribution of m⁶A peaks reads across all mRNAs. g The visual peaks figure of relative abundance of m⁶A sites along SOX2 mRNA in GBM cells. h–i MeRIP-qPCR analysis of intact SOX2 RNA from GBM cells with sh-ALKBH5 transfected or LV-ALKBH5 transfected, *p < 0.05 compared with sh-NC or LV-NC group. j MeRIP-PCR analysis of fragmented SOX2 RNA in GBM cells with or without ALKBH5 knockdown, ****p < 0.0001 compared with sh-NC group. k The qPCR analysis of SOX2 enrichment in chromatin immunoprecipitation, *p < 0.05 compared with the IgG group. l RNA EMSA analysis of ALKBH5 binding to the CpG region of SOX2 gene.