Figure 3.

METTL3 regulated DGCR8-mediated maturation of pri-miR126 in an m⁶A-dependent manner. (A) Interaction between METTL3 and DGCR8 was evaluated by co-immunoprecipitation in HESCs, and immunoglobulin G (IgG) was used as control. Ribonuclease (RNase) was used as indicated. (B) Levels of miRNAs with m⁶A tags in the tissues. NC (n = 10), EU (n = 9), and EC (n = 9). (C and D) Levels of miRNAs were quantified by qRT-PCR in METTL3-overexpressing or METTL3-silenced HESCs. (E–F) Level of pri-miR126 was quantified by qRT-PCR in METTL3-overexpressing or METTL3-silenced HESCs. (G) RNA immunoprecipitation-PCR assays showed obvious enrichment of pri-miR126 bound to DGCR8 in METTL3-overexpressing HESCs. (H) Methylated RIP-PCR assays showed obvious enrichment of pri-miR126 with m⁶A in METTL3-overexpressing HESCs. All experiments were repeated in triplicate or quadruplicate. Data with error bars are presented to indicate the mean ± SEM values. ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001.