BioMed Research International / 2014 / Article / Fig 3

Research Article

Trichostatin A Suppresses EGFR Expression through Induction of MicroRNA-7 in an HDAC-Independent Manner in Lapatinib-Treated Cells

Figure 3

Silence of HDACs did not affect EGFR expression in lapatinib-treated MDA-MB-231 cells. (a)–(c) 231/Lap#6 cells were transfected with control siRNA or specific siRNA against HDAC3 ((a) and (c)) or HDAC7 ((b) and (c)) for 3 days. Total protein lysates prepared from these cells were subjected to Western blot analysis with indicated antibodies ((a) and (c)). Total RNA extracted from these cells was subjected to RT-qPCR. The relative mRNA levels of EGFR, HDAC3, and HDAC7 were normalized to GAPDH expression. (d) Total lysates of 231/Lap# cells transfected with myc-HDAC3 or myc-HDAC7 were subjected to immunoprecipitation with anti-myc antibody. The HDAC activities in the immunoprecipitates were measured in the HDAC activity assays. (e) and (f) 231/Lap#6 cells were transfected with increasing doses of myc-HDAC3 or myc-HDAC7 followed by treatment with TSA. Total lysates were prepared and subjected to Western blot.