Anti actin clone c4

The mouse antibodies used were anti-NAV1 (sc-398641; Santa Cruz Biotechnology; dilution for immunofluorescence [IF]: 1:250, for Western blotting [WB]: 1:1,000), anti-EB1 (610534; BD Biosciences; dilution IF: 1:200), anti-α-tubulin (T6199; Sigma-Aldrich; dilution IF: 1:4,000, dilution WB: 1:10,000), anti-6xHis (552565; BD Biosciences; dilution WB: 1:5,000), and anti-actin (clone C4; MP Biomedicals; dilution WB: 1:10,000). The chicken anti-GFP (GFP-1010; Aves Labs; dilution IF: 1:1,000), the rabbit anti-GFP (TP401; Torrey Pines Biolabs; dilution WB: 1:10,000), and the rat anti-α-tubulin (clone YL1/2; dilution IF: 1:1,000), kindly provided by K. Rogowski (Institut de Génétique Humaine, Montpellier, France), were also used. DyLight Fluor-conjugated secondary antibodies were from Thermo Fisher Scientific, and horseradish peroxidase-conjugated secondary antibodies were form GE Healthcare. ATTO-488–labeled tubulin was kindly provided by D. Portran. Cytochalasin D and blebbistatin (both from Sigma-Aldrich) were added to the cell cultures 20 min before analysis at 1 µM and 40 µM, respectively. SiR-tubulin (Tebu-Bio) was added to cell cultures 90 min before analysis at 50 nM.