Premature centriole disengagement and PCM fragmentation occurs after LIC depletion in HeLa cells. (A, B, and D) Centrin-1–GFP HeLa cells treated with scrambled (control) or LIC 1 and 2 siRNAs were labeled with anti–α-tubulin, anti-PC, and DAPI. (A) Minor multipolar spindles in centrin-1–GFP HeLa cells. White arrowheads, mother centrioles; red arrowheads, daughter centrioles. PC spots without associated centrioles are common at minor poles (cyan arrows) and rare at major poles (yellow arrow). Bars, 5 µm. (B) Centriole number in interphase centrin-1–GFP HeLa cells was scored (>200 cells scored per condition in each of three to four independent experiments, means ± SEM). (C) HeLaM KnD cells were stained with anti-CETN3 to detect centrioles. Centriole number was counted per pole in 100 mitotic cells, in three independent experiments (means ± SEM). (D) Centriole and PC distribution at each major and minor spindle pole in LIC1- and 2-depleted centrin-1–GFP HeLa cells was scored (69 cells, from two independent experiments). ***, P < 0.001, Student’s t test.