Cep131 overexpression promotes centrosome amplification and colon cancer progression by regulating Plk4 stability

a Immunofluorescence analysis of centriole and centrosome amplification in U2OS cells stably expressing HA-Cep131. Cells were stained with anti-Cent (red) and anti-γTub (green) antibodies to visualize centrioles and centrosomes, respectively. Insets are approximately fivefold magnified at the centrosomal region. Scale bar, 10？μm. b Quantification of the proportion of cells with >2 centrosomes or >4 centrioles induced by stably expressing HA-Cep131. c, d Multinucleated cells were induced by HA-Cep131 overexpression in U2OS cells. DNA is visualized in blue. Scale bar, 10？μm. e, f U2OS cells were treated to 2？mM hydroxyurea (HU) for 48？h to induce centrosome amplification and were transfected with siCon or siCep131 duplexes. The graph shows quantification of cells with centrosome amplification (f). Insets are approximately fivefold magnified at the centrosomal region. Scale bar, 10？μm. g Quantification of the proportion of cells with centriole number after treatment of siCon, siCep131, and siPlk4. h Endogenous Cep131 was depleted by siRNA, and cells were transfected with EGFP vector or EGFP-Cep131, containing resistant sequences to siCep131. The graph shows quantification of four-centriole cells. Error bars in b, d, f, g represent means ± SEM from three independent experiments (N？>？300 for each experiment). *P？<？0.05 and **P？<？0.01, unpaired Student’s t tes