Fig. 6

HSPA1A chaperone activity is required for PCNT accumulation at the mitotic centrosome and assembly of mitotic spindles. A Western blot analysis of the expression level of FLAG-tagged wild type and each mutant HSP70. B CGL2 cells stably expressing FLAG-HSP70s were depleted of endogenous HSP70, and then the cells were immunostained with PCNT and α-tubulin to reveal the spindle. The percentages of cells with abnormal spindles were determined by at least 600 mitotic cells from three independent experiments, and Student’s t test was performed. *p < 0.05 comparing FLAG-HSPA1As to pFB-Neo (Neo); ^#p < 0.05 comparing mutants to WT. C Representative images of PCNT (green) and spindles (α-tubulin, red). D Relative PCNT intensity at the spindle pole (SP). Numbers in parenthesis indicate the number of spindle poles examined. Mann–Whitney rank sum test was performed. *p < 0.05 comparing FLAG-HSPA1As to pFB-Neo (Neo); ^#p < 0.05 comparing K71E to WT