Skip to main content
. 2023 Jun 28;13(7):e1318. doi: 10.1002/ctm2.1318

FIGURE 4.

FIGURE 4

CircAFF2 expression is regulated by ALKBH5/YTHDF2‐mediated m6A methylation. Dot blot (A) and m6A RNA methylation assay (B) showing m6A levels in HT‐29 and HCT‐116 cells before and after irradiation. (C) mRNA levels of methylases and demethylases in HT‐29 and HCT‐116 cells following 4 Gy irradiation. (D) qRT‐PCR to detect circAFF2 expression in ALKBH5‐overexpressed or ALKBH5‐depleted HT‐29 and HCT‐116 cells. (E) Methylated RNA immunoprecipitation (Me‐RIP)‐qPCR showed that depletion of ALKBH5 increases the m6A modification of circAFF2. (F) The constructed luciferase reporter plasmids by inserting partial wild‐type circAFF2 sequences or mutated m6A sites (M1–M5) circAFF2 sequences. (G) The luciferase reporters with wild‐type or mutant plasmids were transfected into HT‐29 and HCT‐116 cells (with the perturbation of ALKBH5), followed by the measurement of luciferase activity. (H) qRT‐PCR to detect circAFF2 expression in HT‐29 and HCT‐116 transfected downregulated YTHDF2. (I) qRT‐PCR to detect circAFF2 expression in HT‐29 and HCT‐116 transfected downregulated ALKBH5 or/both downregulated YTHDF2. *** p < .001, ** p < .01, * p < .05.