Figure 2: Sulforaphane enhances expression of acetylated HSP90 and decreases c-myc (a) Level of acetylated HSP90 (Ac-HSP90) was studied in MDA-MB-231 cells treated with sulforaphane (0, 1, 5, 10 and 20 μM) for 24 h. HSP90 was immunoprecipitated (IP) from the tumor cell lysates and immunoblotted (IB) with either anti-HSP90 or anti-acetylated lysine (Ac-lysine) antibody. An increase in expression of acetylated lysines on HSP90 was observed. (b) Level of Ac-HSP90 in MDA-MB-231 cells treated with tubacin (0, 2.5 and 5 μM) for 24 h was studied. HSP90 was immunoprecipitated from the cell lysates and immunoblotted with either anti-HSP90 or anti- Ac-lysine antibody and found to be enhanced. (c) The cells were treated with various concentrations of sulforaphane (0, 1, 5, 10 and 20 μM) for 24 h. 50 μg of isolated protein was electrophoresed on 10% SDS-PAGE gel, blotted and probed with antibody against cmyc. A decrease in c-myc expression was observed. β-actin was used as loading control. (d) Differential expression of c-myc protein after treatment with HDAC6 inhibitor tubacin (0, 2.5 and 5 μM) for 24 h also revealed decrease in c-myc expression. For equal loading of protein, β-actin was used as control.