Figure 2

Analysis interferer efficiency of pGenesil-CENPE. (A)Analysis of CENP-E protein levels by Western blot. Seventy-five micrograms of mitotic extracts derived from LO2 cells treated by nocodazol before detection for 3 h (lane 1-5). (B)shRNA-induced reduction of CENP-E mRNA and protein levels. Reduction of CENP-E mRNA. LO2 cells were transfected with various CENP-E shRNA vectors as indicated, and the mRNA levels were measured 24 h posttransfection by QPCR. Control:negative control (pScramble); mock: transfected Lipofectamine 2000 only alone. (C)Immunofluorescence of CENP-E of LO2 cells 24 h posttransfection with control shRNA vector or CENP-E siRNA. Cells were double stained with DAPI (4,6-diamidino-2-phenylindole) and CENP-E antibodies. Identical exposure times were used for imaging both control and CENP-E shRNA-transfected cells (white arrow point to misaligned chromosome). Bar, 5 μm.