UNK promotes the localization of centriole assembly protein factors. (A) Schematic showing experimental timeline for UNK depletion, S phase arrest, PLK4 overexpression, and fixation for immunofluorescence. (B) Centriole assembly protein localization is reduced in siUNK cells. Left panels: SIM images of RPE-1-Tet-PLK4 Cetn2-GFP p53 WT cells showing fluorescence at centrosomes in UNK-depleted cells with overexpressed PLK4 in S phase. CEP63, CEP192, CEP152, PLK4, and STIL are magenta, and centrioles (centrin) are green. Scale bar, 1.0 μm. Right panels: Mean normalized centrosome fluorescence intensities of centriolar proteins. Graph values are expressed as the means of three and four biological replicates of 25–30 cells per replicate and SD. P values were determined using an unpaired two-tailed t test. (C) UNK depletion promotes ectopic centrin-positive foci. Confocal images of RPE-1-Tet-PLK4 Cetn2-GFP p53 WT cells showing centrin-positive foci in UNK-depleted cells with overexpressed PLK4 in S phase. Centrioles (centrin), grayscale. Arrows indicate sites of ectopic centrin foci. Scale bar, 5 μm. Insets scale bar, 1 μm.