ALKBH5 Exacerbates Aortic Dissection by Promoting Inflammatory Response and Apoptosis of Aortic Smooth Muscle Cells via Regulating lnc-TMPO-AS1/EZH2/IRAK4 Signals in an m6A Modification Manner

Figure 4

ALKBH5 downregulates lncRNA TMPO-AS1 through m6A demethylation in HASMCs. (a) DNA methylation might not be related to the lnc-TMPO-AS1 expression level in HASMCs. (b, c) Histone acetylation had no significant influence on the lnc-TMPO-AS1 level in HASMCs. (d) M6A dot blot assay revealed that overexpression of ALKBH5 markedly reduced m6A level while silencing ALKBH5 increased this level in HASMCs. (e) M6A RIP-qPCR analysis showed 0.3- and 2.5-fold enrichment in m6A antibody levels of lnc-TMPO-AS1 in ALKBH5-overexpressing and ALKBH5 knockdown cells, respectively. (f) The level changes of ALKBH5 had no significant influence on the half-life of lnc-TMPO-AS1. (g, h) Subcellular fractionation analysis indicated that ALKBH5 overexpression could apparently decrease the localization of lnc-TMPO-AS1 to chromatin while ALKBH5 knockdown enhance its accumulation in chromatin. Data represented the from three independent experiments, ; ns represents no statistical difference between groups.

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