Fig. 7.

TFRC serves as a target for METTL3-mediated m⁶A modification via an IGF2BP2-dependent manner. (A) Representative m⁶A dot blot and statistical analysis of m⁶A with or without METTL3 knockdown (n = 3 per group). (B) MeRIP-qPCR analysis showing location of given m⁶A methylation sites in TFRC (n = 4 per group). (C) Decay rate of TFRC mRNA after treatment of actinomycin D (ActD, 5 μg/ml) in METTL3 knockdown HL-1 cells (n = 3 per group). (D) Relative mRNA levels of TFRC in HL-1 cells knocked down with YTH and IGF2BP reader proteins (n = 4 per group). (E) and (F) Relative protein level of TFRC in the absence or presence of IGF2BP2 knockdown (n = 3 per group). (G) Sequences of TFRC RNA fragments with either WT or mutated (MUT) m⁶A sites. (H) RIP assay showing the binding affinity between TFRC mRNA and IGF2BP2 with or without METTL3 knockdown (n = 4 per group). (I) Decay rate of TFRC mRNA following ActD administration in IGF2BP2 knockdown HL-1 cells (n = 3 per group). Mean ± SEM. P values were determined by one-way ANOVA with Tukey's post-hoc test (F, and H) or unpaired two-tailed Student's t-test (A, C, D, and I). P < 0.05, **P < 0.01, and ***P < 0.001.