Fig. 3 | Scientific Reports

Fig. 3

From: ALKBH5 regulates etoposide-induced cellular senescence and osteogenic differentiation in osteoporosis through mediating the m6A modification of VDAC3

Fig. 3

ALKBH5 overexpression inhibited m6A modification of VDAC3. (A) The expression of METTL3, METTL14, FTO, WTAP and ALKBH5 in 293T cells following pcDNA3.1 and their respective overexpressing plasmids transfection was measured by qPCR. (B) The expression of VDAC3 in 293T cells transfected with pcDNA3.1-METTL3, pcDNA3.1-METTL14, pcDNA3.1-FTO, pcDNA3.1-WTAP and pcDNA3.1-ALKBH5 was measured by qPCR. (C) The m6A level of VDAC3 in 293T cells transfected pcDNA3.1 or pcDNA3.1-ALKBH5 was assessed by MeRIP. (D) The enrichment of VDAC3 on ALKBH5 in 293T cells was determined by RIP. (E) The m6A sites were predicted using SRAMP database. (F) The top 3 potential sites in VDAC3. The luciferase activity of (G) wild type and mutant of site 1, (H) wild type and mutant of site 2 and (I) wild type and mutant of site 3 in 293T cells co-transfected with pcDNA3.1 or pcDNA3.1-ALKBH5 was measured by dual luciferase report. (J) qPCR was performed to evaluate VDAC3 expression in BMSCs transfected pcDNA3.1 or ALKBH5 overexpressing plasmids following treatment with 5 µg/mL actinomycin D for 1, 4, 8 and 12 h.

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