Fig. 6: m6A demethylase FTO was involved in the osteoclastogenesis.
a The total m6A methylation level in osteoclast (OC) with obesity-associated protein (FTO) shRNA transfection using Lentiviral vectors was measured (n = 3 independent experiments; mean ± SD). b The cell viability of bone marrow macrophage (BMDM) after treatment with Dac51 (0, 0.5, 1, 2, 4 and 8 μM) was measured (n = 3 independent experiments; mean ± SD). c The total m6A methylation level in OC after treatment with Dac51 (2 μM) was measured (n = 3 independent experiments; mean ± SD). d The relative protein levels of FTO, nuclear factor of activated T cells (NFATc1), matrix metalloprotein 9 (MMP9), and cathepsin K (CTSK) in OC with Lenti-shFTO and Lenti-shNC transfection were determined (n = 3 independent experiments; mean ± SD). e The relative protein levels in OC after treatment with Dac51 were determined (n = 3 independent experiments; mean ± SD). f The representative TRAP staining images of OC were shown after transfecting with Lenti-shFTO and Lenti-shNC for 4 days, respectively (n = 3 independent experiments). g The representative TRAP staining images of OC after treatment with Dac51 for 4 days at different concentrations were shown (n = 3 independent experiments). h The bone resorption pits on bone slices after transfecting with Lenti-shFTO as compared to Lenti-shNC for 4 days were stained by toluidine blue. The arrowheads marked in yellow color indicate the bone resorption pits (n = 3 independent experiments). Scale bar indicates 100 μm. i The bone resorption pits on bone slices after treatment with Dac51 for 4 days were stained by toluidine blue (n = 3 independent experiments). Arrowheads indicate the bone resorption pits. The Lenti-shNC group BMDM cells were received the treatment of unedited plasmid carried by lentiviral vector. The major statistical procedures applied were: Shapro–Wiik test (a–e), F-test (b, d, e), Student-t test (a, c), One-way ANOVA and Dunnett-t test (d, e), Kruskal–Wallis H test and Dunnett-t test (d, e). The statistical test used was two-sided. Source data are available online for this figure.
