Fig. 5.

HOXC10 directly binds to NOD1 promoter and promotes its expression. a HOXC10 regulated the NOD1 promoter activity. H441-BM cells were then harvested for a luciferase assay. b Electrophoretic mobility shift assay. The sequence of HOXC10 containing NOD1 promoter binding site (Left) using the JASPAR database. The WT and Mut probes of NOD1 are shown (left). The mutated site is indicted in red. The binding complex of indicated probes and HOXC10 protein was indicated by arrows (Right). c ChIP assay showing the in vitro binding of HOXC10 and NOD1 promoter. d, e Ectopically expressed NOD1 restored the clonogenic growth of HOXC10-depleted cells. The relative protein levels (d), and colony-formation ability (e) of indicated cells transfected with the indicated siRNAs. f–i The relative protein levels (g) Scale bars, 50 μm, migration ability (i), osteoclast differentiation ability (h) Scale bars, 100 μm, and MMP9 mRNA levels (j) of indicated cells transfected with the indicated siRNAs. Data in (a, e, g, h, i) represent the mean ± s.e.m. of three technical replicates, representative of three independent experiments with similar results. Statistical analysis in (a, e, g, h, i) were performed one-way ANOVA with Tukey’s multiple comparison test, and in (c) performed unpaired two-sided Student’s t test, *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant