Fig. 6.

STAT3 inhibitor synergize with HOXC10 inhibition in vitro and in vivo. a The top 5 altered pathways of the RNA-seq data. The RNA samples extracted from HOXC10 knockdown bone metastasis tissues after intracardiac injection at 7 and 28 days (n = 3). b qPCR validation of IL6 mRNA level. c Protein expression levels. Bone metastasis tumors were harvested at day 7, and day 28 in Fig. 3d. Two biologically independent samples per group from five independent samples are shown. d, e IL-6 treatment reduced HOXC10 knockdown-mediated cytotoxicity. Cells were treated with IL-6 at the concentration of 20 ng/mL. The relative clonogenic viability (d) and migration ability (e) were normalized to vehicle-treated control. Scale bars, 50 μm. f, g TTI-101 treatment increased HOXC10 knockdown-mediated cytotoxicity. Cells were treated with TTI-101 at the concentration of 4 μmol/L. The relative clonogenic viability (f) and migration ability (g) were normalized to vehicle-treated control. Scale bars, 50 μm. h The osteoclast differentiation ability of HOXC10 inhibition plus TTI-001 (4 μmol/L) was assessed. Scale bars, 100 μm. i Representative IVIS bioluminescence imaging of indicated mice at day 35. After 7 days intracardiac injection, mice were treated with TTI-101 (25 mg/kg) for an additional 28 days (n = 6 per group). j The growth of bone metastasis in (i) was quantified (n = 6 per group). k Representative micro-CT images of spine. Red arrows, osteolytic lesions. Bar graph of quantitative micro-CT analysis of osteolytic lesions of spine from the moribund mice (n = 3 per group). Scale bars, 1 mm. l Survival curves for indicated mice (n = 6 per group). Data in (b, d, e, f, g, h) represent three technical replicates, representative of three independent experiments with similar results. Data in (b, d, e, f, g, h, j) shown as mean ± s.e.m. Panels (b, d, e, f, g, h, k) were performed one-way ANOVA with Tukey’s multiple comparison test, (j) performed two-way ANOVA with Sidak’s multiple comparisons test, and (l) performed log-rank test, *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant