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 5

lncRNA TMPO-AS1 exhibits its biological roles in HASMCs via regulating IRAK4. (a) The expression levels of a large number of genes changed significantly after the lnc-TMPO-AS1 knockdown. (b) IRAK4 had the most dramatic expression change when lnc-TMPO-AS1 was overexpressed. (c) The protein level of IRAK4 was negatively regulated by lnc-TMPO-AS1. (d) The result of subcellular fractionation analyses indicated that lnc-TMPO-AS1 was predominantly located in the nucleus. (e) RIP assay revealed that lnc-TMPO-AS1 can combine with HuR, LSD1, and EZH2, with the strongest binding to EZH2. (f) The signals of lnc-TMPO-AS1 and EZH2 were confirmed to be colocalized in HASMCs by FISH and immunofluorescence. (g, h) No significant changes in EZH2 expression appeared in both pcDNA-TMPO-AS1 and si-TMPO-AS1 cells compared to the negative control. (i) EZH2 was downregulated in the AngII treatment group. (j, k) The expression of EZH2 was also reduced in aortas from AD patients and exhibited a positive correlation with lnc-TMPO-AS1 expression (, ). (l, m) EZH2 knockdown significantly enhanced the mRNA and protein levels of IRAK4. (n) ChIP assays demonstrated that lnc-TMPO-AS1 knockdown reduced the binding of EZH2 and H3K27Me3 levels at the promoters of IRAK4. Data represented the from three independent experiments; , .

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