Fig. 3

HSP70 is required for the 3D assembly of CEP215, PCNT and γ-tubulin at the mitotic centrosome. A 2D projections of the averaged 3D-SIM images of each indicated centrosome marker in control interphase (a) and mitotic cells (c) and HSP70-depleted interphase (b) and mitotic (d) cells. Merged images show the relative distribution of each centrosome marker with respect to γ-tubulin. B, C Box plots show the relative diameter and volume of each marker in control and HSP70-depleted mitotic cells with the number of samples (n) indicated. Mann–Whitney rank sum test was performed. *p < 0.05 comparing shHSP70 to pLKO.1. D Surface-rendered images of the centrosome with or without HSP70. E (a) Representative wide-field image of a HSP70-depleted mitotic cell with multipolar spindle. Two poles with centriole marker (CEP164) are indicated with red boxes, and the other extra spindle poles without CEP164 are indicated by white boxes. (b) Averaged SIM images of the extra spindle poles