METTL3 Promotes Osteo/Odontogenic Differentiation of Stem Cells by Inhibiting miR-196b-5p Maturation

Figure 4

METTL3-dependent m6A methylation regulates miR-196b-5p maturation by the microprocessor protein DGCR8. (a) Co-IP assay showed that the METTL3 interacted with DGCR8 by western blot in SCAPs. (b, c) Co-IP of DGCR8, METTL3, and associated RNAs from METTL3 overexpression cells and METTL3 knockdown cells. Western blot using the DGCR8 and METTL3 antibodies and IgG was used as the control for the IP. (d, e) The expression of pri-miR-196b was verified by qRT-PCR in METTL3 overexpression and knockdown cells. (f, g) The expression of miR-196b-5p was verified by qRT-PCR in METTL3 overexpression and knockdown cells, respectively. (h) The detection of pri-miR-196b m6A modification level by MeRIP in control or METTL3 overexpression cells followed by qRT-PCR. (i) Detection of pri-miR-196b binding to DGCR8 by immunoprecipitation of DGCR8-associated RNA from control and METTL3 overexpression cells followed by qRT-PCR. U6 was used as an internal control for pri-miR-196b and miR-196b-5p. Student’s -test was used to analyze the statistical significance. All error bars signify standard deviations (). and .

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