Research Paper Volume 13, Issue 17 pp 21134—21141

RNA N6-methyladenosine demethylase FTO promotes osteoporosis through demethylating Runx2 mRNA and inhibiting osteogenic differentiation

FTO represses the expression of Runx2 through its m6A demethylase activity. (A) The methylated RNA (m6A) level of Runx2, Sp7, Alp and Bglap in normal and FTO over-expressed BMSCs. *PB) The mRNA level of Runx2 in Wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *PC) The protein level of Runx2 in wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *PD) The methylated RNA (m6A) level of Runx2 in FTO down-regulated, wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *PE) Correlation between the mRNA level of FTO and Runx2 in the 20 cases of OP patients (R2=0.3735, PF) Half-life of Runx2 mRNA in FTO down-regulated, wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs after transcription inhibition (TI). All values were normalized to 18S rRNA.

Figure 3. FTO represses the expression of Runx2 through its m6A demethylase activity. (A) The methylated RNA (m6A) level of Runx2, Sp7, Alp and Bglap in normal and FTO over-expressed BMSCs. *P<0.05. (B) The mRNA level of Runx2 in Wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *P<0.05. (C) The protein level of Runx2 in wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *P<0.05. (D) The methylated RNA (m6A) level of Runx2 in FTO down-regulated, wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs. *P<0.05. (E) Correlation between the mRNA level of FTO and Runx2 in the 20 cases of OP patients (R2=0.3735, P<0.01, Pearson’s correlation test). (F) Half-life of Runx2 mRNA in FTO down-regulated, wild-type or demethylase mutated FTO (R96Q) over-expressed BMSCs after transcription inhibition (TI). All values were normalized to 18S rRNA.