Fig. 2

miR-6359 is enriched in VPA-treated osteoclast precursors and is responsible for osteoclast differentiation. a Representative images displaying the TRAP-positive multinuclear cells (n = 3). Scar bar: 100 μm. b Quantitative analysis of TRAP-positive area/total area (%). c Western blot analysis of the impact of VPA treatment on Nfatc1, Ctsk and Trap protein levels. d TRAP staining of osteoclast precursors treated with VPA at different stages of osteoclastogenesis (n = 3). Scar bar: 100 μm. e The area of TRAP-positive cells/total area (%) was counted. f Volcano plot showing genes or miRNAs with a cut-off fold-change of ≥ 4 or ≤ −4 and a p value of < 0.01. g qRT-PCR detection of the relative abundance of miR-6359 in osteoclast precursors treated with or without VPA (n = 4). h Relative abundance of miR-6359 in the serum of mice treated with or without VPA, as determined by qRT-PCR analysis (n = 15). i qRT-PCR detection of miR-6359 expression level in various tissues, such as heart, liver, spleen, lung, kidney and bone following VPA treatment (n = 4, 5 or 6). j, k CX3CR1⁺, Csf1R⁺ osteoclast precursors were identified (j) and the miR-6359 level was measured through qRT-PCR analysis (k) (n = 6). l miR-6359-FISH and immunofluorescence for LY6G, CD11b, CD3, CD90 and CX3CR1 were performed. Scar bar: 5 μm. Data are presented as the mean ± SEM (n ≥ 3) (*p < 0.05; **p < 0.01; ***p < 0.001)