Fig. 3. METTL3 silencing reduced m⁶A modification in OS cells and thereby suppressed HDAC5 expression.

A, B m⁶A level in 50 pairs of OS tissues and adjacent tissues was detected by m⁶A quantitative analysis. B m⁶A level in human OS cell lines (U2OS, HOS, SAOS2) and normal osteoblasts (hFOB1.1) was detected by m⁶A quantitative analysis. E, F HDAC5 expression in human OS cell lines (U2OS, HOS, SAOS2) and normal osteoblasts (hFOB1.1) was determined using RT-qPCR and western blot. G The correlation between HDAC5 and METTL3 expression in OS tissues was analyzed using Pearson correlation analysis. H The m⁶A modification level of HDAC5 mRNA in OS cells after intervention with METTL3 expression was detected using MeRIP-qPCR. I–K HDAC5 expression and stability in OS cells after intervention with METTL3 expression was measured using RT-qPCR and western blot. N = 50. The cell experiment was repeated three times independently. Data in panels A, B/D–F/H–K are presented as mean ± standard deviation. Data in panel C were analyzed using paired t-test and data in panels A/D were analyzed using t-test. Data in panels B/E–F/H–J were analyzed using one-way ANOVA, and data in panel K were analyzed using two-way ANOVA, followed by Tukey’s multiple comparisons test, *p < 0.05, **p < 0.01.