Fig. 6

ERVWE1 reduced the global m6A level dependent ALKBH5 contributed to HTR1B upregulation. A, B The global m6A level was measured by A dot blot, and B the EpiQuik m6A RNA Methylation Quantification Kit (Colorimetric) in ERVWE1 expressed groups in SH-SY5Y cells. Methylene blue (MB) was leveraged as a control. C The relative mRNA levels of methyltransferase (METTL3 and METTL14), demethylases (ALKBH5 and FTO), and reader proteins (YTHDC1, YTHDC2, YTHDF1, YTHDF2, and YTHDF3) in the control and ERVWE1 groups by RT-qPCR in SH-SY5Y cells. D The western blot results of METTL3, METTL14, ALKBH5, FTO, and YTHDC2 in SH-SY5Y cells. E ALKBH5 promoter activity as measured by Promega Dual-Luciferase Reporter Assay. F, G The global m6A levels were measured by dot blot and the EpiQuik m6A RNA Methylation Quantification Kit after co-transfection with ERVWE1 (or empty plasmid) and siALKBH5 (or siNC) in SH-SY5Y cells. H m6A level of HTR1B was determined by Me-RIP-qPCR assay after overexpression of ERVWE1 in SH-SY5Y cells. I ERVWE1 increased the enrichment of ALKBH5 in HTR1B mRNA by RIP-qPCR analysis in SH-SY5Y cells. J The expression levels of HTR1B and downstream proteins were detected by western blot when ERVWE1 was co-transfected with siALKBH5 or siNC in SH-SY5Y cells. Data are mean ± SD. Statistical analysis: Student’s t-test (two groups) and one-way ANOVA (three groups) (^ns p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001). All experiments were repeated 3 times