Effects of p27Kip1- and p53- shRNAs on kanamycin damaged mouse cochlea

Figure 1 Short hairpin RNA testing in COS cells. A: Western blot analysis with anti-p27^Kip1 antibody. Proteins were extracted from untransfected COS cells and COS cells transfected with Bluescript (pBS), p27^Kip1 cDNA (p27-cDNA) or p27^Kip1 cDNA together with 4 different p27^Kip1 short hairpin RNA (shRNA) constructs, p27(1-4). p27^Kip1 specific bands are marked on the right side. A decrease in p27^Kip1 level was observed with p27(2), p27(3) and p27(2+3) after 24 h of transfection; B: Western blot analysis with anti-p53 antibody. Proteins were extracted from untransfected COS cells (COS), p53 null cells (p53-/-), p53 heterozygous null cells (p53+/-), COS cells transfected with p53 shRNA. Cells were collected 24 h or 48 h after transfection. p53 specific bands are marked on the right side. A decrease in p53 level was detected after 24 h of transfection. Kaleidoscope molecular weight (MW) standards are marked on the left side. MEF: Mouse embryonic fibroblasts.