Figure 4

Effect of lactacystin on TSA induced cyclin D1 degradation. A, MCF-7 cells were pretreated with lactacystin (10 μM) for 1 h and then treated with TSA (1 μM) for 6 h. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against cyclin D1 and actin. B, effect of ammonium chloride and lactacystin on TSA induced cyclin D1 degradation. MCF-7 cells were pretreated with the indicated concentrations of ammonium chloride (10 mM) and/or lactacystin (10 μM) for 1 h and then treated with TSA (1 μM) for 6 h. Immunoblot analysis was done with antibodies against cyclin D1 and actin. C, Effects of ALLM, lactacystin and LMB on TSA induced cyclin D1 degradation. MCF-7 cells were pretreated with ALLM (100 μM), lactacystin (10 μM) or LMB (10 ng/mL) for 1 h and then treated with TSA (1 μM) for 6 h. Immunoblot analysis was done with antibodies against cyclin D1 and actin. D, Lactacystin and ALLM induced cyclin D1 loss is inhibited by MG132. MCF-7 cells were treated with lactacystin (10 μM) or ALLM (100 μM) with or without MG132 (50 μM) for 6 h. Immunoblot analysis was done with antibodies against cyclin D1 and actin.