Figure 2.

BITC treatment decreases sRANKL expression and secretion. (A) Quantitation of intracellular sRANKL level in MDA-MB-231 (MDA-231) cells after 24- or 48-h treatment with DMSO or the indicated doses of BITC. (B) Quantitation of sRANKL secretion in CM of MDA-MB-231 (MDA-231) cells after 24- or 48-h treatment with DMSO or the indicated doses of BITC. (C) Quantitation of sRANKL secretion in CM of RAW264.7 (RAW) and MDA-MB-231 (MDA-231) co-culture after 6 days of treatment with DMSO or the indicated doses of BITC. For panels (A–C), columns indicate average of triplicate samples from a representative experiment, and bars indicate standard deviation. P value relative to respective DMSO-treated control was determined by one-way analysis of variance (ANOVA) with Dunnett’s adjustment. (D) Western blotting for sRANKL, OPG and GAPDH proteins using whole cell lysates from MDA-MB-231, MCF-7 and SK-BR-3 cells after 24- or 48-h treatment with DMSO or the indicated doses of BITC. The numbers above bands represent fold changes of proteins relative to corresponding DMSO-treated control. (E) Ratios of quantitative values of sRANKL/OPG from western blotting. Columns indicate average of three different experiments, and bars indicate standard deviation. P value relative to respective DMSO-treated control was determined by one-way ANOVA with Dunnett’s adjustment. Each experiment was repeated at least three times and the results were consistent.