Research Paper Volume 13, Issue 19 pp 23361—23375

piRNA-36741 regulates BMP2-mediated osteoblast differentiation via METTL3 controlled m6A modification

Silencing METTL3 impeded osteogenic differentiation of BMSCs. BMSCs were infected with Lv-sh-NC or Lv-sh-METTL3, and then cultured in osteogenic differentiation medium for 14 days. (A) The expression of piR-36741 was measured on day 14. (B) The global m6A level of BMSCs with or without METTL3 knockdown was analyzed with the EpiQuik™ m6A RNA methylation quantification kit. (C) The mRNA levels of METTL3, RUNX2, COL1A1, OCN and OPN were detected on day 14. (D) Images of ALP staining on day 14 (100×). (E) ALP activity was determined on day 14. (F) Images of ARS staining on day 14 (100×). (G) Quantitative analysis of ARS accumulation on day 14. N = 5 in each group. *P **P

Figure 4. Silencing METTL3 impeded osteogenic differentiation of BMSCs. BMSCs were infected with Lv-sh-NC or Lv-sh-METTL3, and then cultured in osteogenic differentiation medium for 14 days. (A) The expression of piR-36741 was measured on day 14. (B) The global m6A level of BMSCs with or without METTL3 knockdown was analyzed with the EpiQuik™ m6A RNA methylation quantification kit. (C) The mRNA levels of METTL3, RUNX2, COL1A1, OCN and OPN were detected on day 14. (D) Images of ALP staining on day 14 (100×). (E) ALP activity was determined on day 14. (F) Images of ARS staining on day 14 (100×). (G) Quantitative analysis of ARS accumulation on day 14. N = 5 in each group. *P < 0.05, **P < 0.01. Each test was independently repeated at least three times.