Rabbit anti flag

Immunoblot analysis was performed as described previously (12 ). Samples were subjected to SDS-PAGE followed by western blotting. After blocking with 10% skim milk, blots were incubated with primary antibodies overnight at 4°C and then incubated with secondary antibodies conjugated to horseradish peroxidase (1:10,000, # 805-035-180, # 111-036-045, or # 115-036-062, Jackson ImmunoResearch, PA, USA) for 2 hours at room temperature. Antibodies used: goat anti-KIF9 1:100 (#SC99958, Santa Cruz Biotechnology, CA, USA); rabbit anti-ACTB 1:1000 (#PM053, Medical & Biological Laboratories, Aichi, Japan); goat anti-BASIGIN 1:500 (#SC9757, Santa Cruz Biotechnology), mouse anti-acetylated tubulin 1:1,000 (#T7451, Sigma-Aldrich, MO, USA); mouse anti-AKAP4 1:5,000 (#611564, BD Biosciences, CA, USA); mouse anti-phosphotyrosine 1:1000 (#05-321, Merck Millipore, MA, USA); rabbit anti-RSPH9 1:200 (#HPA031703, Atlas Antibodies); rat anti-PA 1:1000 (#012-25863, FUJIFILM Wako Pure Chemical, Osaka, Japan); and rabbit anti-FLAG 1:1000 (#PM020, Medical & Biological Laboratories). Immunoreactive proteins were detected by using an ECL western blotting detection kit (GE Healthcare, Little Chalfont, UK).