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Fig. 4 | Cell Division

Fig. 4

From: HSP70 is required for the proper assembly of pericentriolar material and function of mitotic centrosomes

Fig. 4

HSP70 is required for the interaction between PCNT and CEP215. A GSD images of PCNT (green) and CEP215 (red) in control (a) and HSP70-depleted (d) mitotic cells. Lines of different directions across the center are shown in merged images. (b, c, e and f) The intensities along each line within a central 1.8-μm region were plotted. At least 20 spindle poles were analyzed and five lines were drawn in each pole. Mean distribution curves are shown in green for PCNT and red for CEP215. The distance between two peak regions (mean ± SD) and the peak-center intensity ratio (median), which were determined as described, are presented and were tested by Student’s t test and Mann–Whitney rank sum test, respectively. *p < 0.05 comparing shHSP70 to pLKO.1. B PLA was performed as described in “Methods” to measure the interaction between PCNT and CEP215. Left, representative images of PLA signal generated using heat-inactivated (−) or normal activity (+) PLA reagents. Right, representative confocal images of cells depleted of HSP70 or treated with 40 μM PES 30 min before fixation are shown. C Relative PCNT and CEP215 PLA intensity at the spindle pole with the number of examined spindle poles (n) indicated. Mann–Whitney rank sum test was performed. *p < 0.05 comparing shHSP70 to pLKO.1 or PES to untreated. D Representative GSD images of colocalization between PCNT (green) and CEP215 (red) at the spindle pole of mitotic cells from control or HSP70-depleted cultures. E The level of PCNT and CEP215 colocalization was measured by Pearson’s correlation coefficient, and the coefficient ranking is shown with the number of examined spindle poles (n) indicated. Mann–Whitney rank sum test was performed. *p < 0.05 comparing shHSP70 to pLKO.1

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