Fig. 7
ERVWE1 upregulated HTR1B via enhancing its mRNA stability. A–D The mRNA half-life (t1/2) of HTR1B in SH-SY5Y cells after transfection with A ERVWE1, B ALKBH5, C siALKBH5 or siCtrl, and co-transfection with D ERVWE1 (or control) and siALKBH5(or siNC) by Actinomycin D (5 µg/mL) treatment. Cells were harvested at 0, 3, and 6 h. The expression level was normalized to that at “0 h”, and GAPDH was used as a control. The mRNA decay rate was analyzed by nonlinear regression curve fitting (one-phase decay model). E Putative m6A modification sites in the coding sequence of HTR1B and synonymous mutations. F, G Relative luciferase activity of HTR1B with either wild-type or mutant m6A sites after co-transfection with ERVWE1 or control vector (F) and tri-transfection with ERVWE1 (or control) and siALKBH5 (or siNC) vectors (G) in SH-SY5Y cells. H The decay of HTR1B mRNA in SH-SY5Y cells with CDS-WT and CDS-mutant. I–L Decay of HTR1B mRNA with ERVWE1 overexpression versus control in the HTR1B-CDS-WT and mutant m6A sites groups in SH-SY5Y cells. M The protein level of HTR1B after mutation of m6A sites in SH-SY5Y cells. N The protein level of HTR1B between pcDNA3.1 and pcDNA3.1-ERVWE1 group after transfected with mutation plasmids in the SH-SY5Y. Data are mean ± SD. Statistical analysis: Student’s t-test or one-way ANOVA (ns p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001). All experiments were repeated 3 times