Fig. 2. Localization of Cep131 at the centriole.

Immunofluorescence analysis of direct localization of Cep131 at the centriole. U2OS cells were co-stained for Cep131 (red) with centriolar marker proteins, such as anti-Cent a and Plk4 b. Scale bar, 0.5μm. c Triple staining of U2OS cells with antibodies against anti-Cent (blue), Plk4 (green), and Cep131 (red). Schematic illustration of the localization of Cep131 at the outer wall of centriole (right panel). Scale bar, 0.5 μm. d Asymmetrical localization of Cep131 at the mother centriole. Cep131 co-stained with Cent (far-red) and mother centriole-enriched protein, hCenexin (green). Schematic illustration to recognize each centriole, mother (M) and daughter (D) centriole. Scale bar, 0.5 μm. e Quantification of fluorescence intensity of Cep131 at each centriole (mother and daughter). Around 50 centrioles from three independent experiments were measured for each condition. ***P < 0.001, unpaired Student’s t test. f, g U2OS cells were treated with shGL2 (control) or shPCNT duplexes and were fixed by two different methods, common fixation (No extraction) and signal extraction (Extraction), to verify the localization of Cep131 at centriolar satellites and the centriole, respectively. Around 50 cells from three independent experiments were measured for each condition. ***P < 0.001, unpaired Student’s t test