Pharicin A induces chromosome congression defect and aberrant BubR1 localization. (A) HeLa cells treated with pharicin A (8 µM) or transfected with CENP-E, CENP-F or KIF18A siRNAs for 24 h were fixed and stained with antibodies to α-tubulin (green) and p-H3S10 (red). DNA was stained with DAPI (blue). Representative images were shown. (B) Percent of cells with lagging/unaligned chromosomes as shown in A were summarized from 300 mitotic cells treated with pharicin A or transfected with CENP-E, CENP-F or KIF18A siRNAs for 24 h. (C) HeLa cells treated with pharicin A (8 µM) or paclitaxel (Taxol, 10 nM) or transfected with Sgo1, CENP-E or KIF18A siRNAs for 24 h were collected and lysed. Equal amounts of cell lysates were blotted for CENPE , KIF18A, BubR1 or β-actin. (D) HeLa cells treated with vehicle or pharicin A (8 µM), and/or transfected with KIF18A or CENP-E siRNAs for 24 h were fixed and stained with antibodies to BubR1 (Red) and β-tubulin (Green). DNA was stained with DAPI (blue). Each experiment was repeated for at least three times. Representative images were shown.