Sc 81431

Cells were fixed with 1.5% formaldehyde in PBS for 5 min and post-fixed with ice-cold methanol for 5 min at −20 °C. After several washes in PBS, the cells were blocked in 1% BSA (Sigma) and 0.05% Tween-20 (Sigma) in PBS for 15–0 min and then incubated with primary antibodies. Alexa Fluor 555-, 405-, 488- or 647-conjugated secondary antibodies (Invitrogen) were used to visualize the proteins. DNA was stained with Hoechst 33342 (Invitrogen). The following primary antibodies were used for immunostaining in this study: mouse monoclonal anti-Sas6 (1:500; sc-81431; Santa Cruz), mouse monoclonal anti-γ-tubulin (1:5,000; T6557; Sigma), mouse anti-centrobin (1:600; ab70448; Abcam), rabbit anti-Cep135 (1:500; ab75005; Abcam), rabbit anti-Cep152 (1:4,000; A302-479 A; Bethyl), rabbit anti-γ-tubulin (1:5,000; T3559; Sigma), rabbit anti-Cep215 (1:2,000; 06-1398; Millipore), goat anti-CPAP (1:500; sc-66747; Santacruz), rabbit anti-CPAP (1:500; 11517-1-AP; Proteintech), mouse anti-GT335 (1:2,000; AG-20B-0020-C100; Adipogen), mouse anti-Plk1(pT210) (1:500, 558400; BD Biosciences) rabbit anti-Plk4 (1:100; kind gift from Dr Kyung S. Lee, NIH Bethesda, USA), rabbit anti-hPOC5 (1:1,000; a kind gift from J. Azimzadeh, Institute Jacques Monod, Paris, France), rabbit anti-Cep120 (1:1,000; a kind gift from Moe R Mahjoub, Washington University in St Louis, USA).