Figure 1: Sulforaphane down-regulates HDAC6 in breast cancer cells (a) Constitutive expression of HDAC6 in normal breast epithelial cells MCF-12F and metastatic breast cancer cells MDAMB- 231 as obtained from western blot analysis revealed a higher expression of protein in cancer cells. β-actin was used as loading control. (b) Cells were exposed to various concentrations of sulforaphane (0, 1, 5, 10 and 20 μM) for 24 h. HDAC6 was analyzed by western blot analysis using specific antibody and was found to be down-regulated by sulforaphane in a dose dependent manner. β- actin was used as control to ensure equal loading of protein. (c) Cells were incubated with sulforaphane (0, 1, 5, 10, and 20 μM) for 24 h to assess mRNA levels of HDAC6. Total RNA was isolated and reverse transcribed. The resulting cDNAs were subjected to PCR with primers and the reaction products were subjected to electrophoresis using 2% agarose gel and visualized by EtBr staining. A decreasing trend at mRNA level was observed after treatment with increasing concentrations of sulforaphane. GAPDH was used as an internal control.