Fig. 3. Mislocalization of Plk1 in CHO1/MKLP-1-depleted cells during late mitosis. (A,B) HeLa cells on coverslips were first synchronized at late G1 phase with double-thymidine block. Upon release from the second thymidine block, cells were directly transfected with synthetic 21-nucleotide double-stranded RNA targeting CHO1/MKLP-1. Cells were fixed 10 hours later and subjected to double staining with anti-CHO1 and anti-Plk1 antibodies. DNA was stained with DAPI. (C,D) HeLa cells were transfected withpBS/U6-GFP-Plk1. 48 hours after transfection, cells were harvested and subjected to anti-Plk1 western blot (C) or anti-CHO1 staining (D). Scale bar, 10 µm.