Figure 8.

YTHDC2 regulates the stability of CYLD through m⁶A modification. (A) Prediction score of m⁶A distribution in CYLD sequence according to the sequence-based RNA adenosine methylation site predictor online tool. (B) meRIP-quantitative PCR and (C) meRIP-PCR results showed amplification of sites 2 to 5, indicating m⁶A modification in these four segments. Anti-IgG antibody was used as control. (D and E) Western blot results showing the relative protein expression level of CYLD in METTL3, METTL14, FTO and ALKBH5 knocked down H1299 cells. (F and G) Western blot result showing the relative protein expression level of CYLD in H1299 cells treated with DAA, a global methylation inhibitor, and with meclofenamic acid, a FTO inhibitor. (H and I) Western blot results showing the relative protein expression level of CYLD in YTHDC2-overexpressing H1299 cells treated with or without 3-DAA. YTHDC2, YTH domain containing 2; CYLD, cylindromatosis; meRIP, m⁶A methylated RNA immunoprecipitation; m⁶A, N6-methyladenosine; METTL, methyltransferase-like; FTO, fat mass and obesity-associated protein; DAA, 3-deazaadenosine.