Mouse anti β catenin

Cells were fixed with 4 % paraformaldehyde in PBS for 10 min at room temperature and then permeabilized with 0.5% Triton™ X-100. For staining MTs and dynein/dynactin, cells were fixed in -20 °C methanol for 5 min; for dynein/dynactin s taining, starved cells were additionally pretreated for 1 hr with 10 μM nocodazole before fixation [65 (link)]. Cells were blocked with blocking buffer (1× PBS; 5% BSA; 0.3% Triton™ X-100) in room temperature for 1 hr and then incubated with primary antibodies diluted in the antibody buffer (1X PBS; 1% BSA; 0.3% Triton™ X-100) for 1 hr at room temperature or overnight at 4 °C. The following antibodies were used: rabbit anti-nesprin 2G [11 (link)] (1:100), rabbit anti-pericentrin (1:400; Covance), mouse anti-pericentrin (1:100; BD biosciences), mouse anti-β-catenin (1:400; Thermo Fisher Scientific) rat anti-Tyr tubulin (1:40; European Collection of Animal Cell Cultures), mouse anti-myc (1:400; Roche), chicken anti-GFP (1:100; EMD Millipore). Rhodamine-phalloidin (1:200; Thermo Fisher Scientific) was used to stain F-actin. After washing with PBS, cells were incubated with fluorescent secondary antibodies absorbed to minimize species cross-reactivity (Jackson ImmunoResearch) for 1 hr at room temperature and mounted in Fluoromount-G (Southern Biotech).