HeLa and DLD1 cells differ in their mechanism of acentriolar spindle assembly. (A) Immunofluorescence of control DMSO- versus centrinone-treated HeLa and DLD1 cells. Scale bar, 5 µm. (B) Immunoblots of knockout HeLa and DLD1 cells. Two independent clones are shown for each line. α-tubulin is a loading control. (C) Immunofluorescence of control and one of the knockout clones per target gene in HeLa and DLD1 cells. Scale bars, 5 µm. (D) Mitotic outcomes in live imaging of chromosome dynamics for the indicated conditions. Protocol is schematized on the top left. n = 50 cells per condition. (E) Microtubule imaging in mitotic cells for the indicated conditions. Protocol is schematized on the top left. n = 50 cells per condition. Scale bar, 5 µm. C, centrinone; D, DMSO; tub, tubulin.