Fig. 7
FTO-mediated m6A modification in BMSCs regulates sclerostin stability. A M6A-Seq identified the m6A site in CDS-3’-UTR junction region of SOST transcript. B M6A consensus motif and metagene analysis of the SOST transcript. C SRAMP prediction results of m6A sites on SOST transcript. D MC3T3 cells were stably infected with FTO-shRNA or negative control-shRNA by lentivirus vector, the m6A modification of SOST transcripts were detected by m6A immunoprecipitation (MeRIP)-qPCR (n = 3). E PRIdictor Database displays the potential Protein-RNA binding sites of FTO and YTHDF2 on SOST transcript. F RPISeq Prediction estimates the Interaction probability of FTO and YTHDF2 protein to SOST transcript. G RNA immunoprecipitation-qPCR assay evidenced the SOST transcript enrichment precipitated by anti-FTO antibody in BMSCs treated with or without AGEs. IgG acted as the blank control (n = 3). H RNA immunoprecipitation-qPCR assay evidenced the SOST transcript enrichment precipitated by anti-YTHDF2 antibody in MC3T3 cells stably infected with LV-shNC or LV-shFTO. IgG acted as blank control (n = 3). I RNA stability assay showed the half-life (t1/2) of SOST mRNA in BMSCs treated with AGEs exposure and FTO knockdown (n = 3). J The relative level of SOST transcripts in subcellular fractions of BMSCs infected with LV-shNC or LV-shFTO was detected by RT-qPCR. β-actin and U6 were employed as cytoplasmic and nuclear loading controls, respectively (n = 3). Data are expressed as the mean ± SEM. ns, not significant. *P < 0.05. **P < 0.01. ***P < 0.001