Figure 3.

VIRMA knockdown in PC-3 cells caused global m⁶A reduction. (A) Sanger sequencing of PC-3 Wild Type (control) and CRISPR edited sequences in the regions around the guide sequence (the horizontal black underlined region represents the RNA guide sequence, the horizontal red underline is the PAM site and the vertical black dotted line represents the cut site); (B) Western blot quantification of the total amount of METTL3, METTL14, WTAP, and VIRMA in the nuclear protein fraction after VIRMA knockdown. Relative protein abundance was determined by normalization with LAMINB1. Bars represent mean ± SD based on 3 independent experiments, * p < 0.05, Mann–Whitney U test. (C) Global m⁶A modification levels in total RNA after VIRMA silencing in PC-3 cells, * p < 0.05 and ** p < 0.001, Mann–Whitney U test; (D) VIRMA expression and m⁶A levels assessed by confocal immunofluorescent assay (scale bar = 20 µm).