Clip170

The following antibodies were commercially purchased and used at the indicated dilutions for immunofluorescence (IF) and immunoblotting (IB); Actin (I-19, Santa Cruz Biotechnology, IB; 1/2000), α-tubulin (B-5-1-2, Sigma, IF; 1/1000), BubR1 (Bethyl Laboratories, IF; 1/500), CENP-E (Sigma, IF; 1/500, IB; 1/2000), CENP-F (Abcam, IF; 1/1000), CLIP170 (Santa Cruz Biotechnology, IF; 1/500), dynein intermediate chain (DIC, Sigma, IF; 1/500), Hec1 (9G3, Abcam, IF; 1/1000, IB; 1/2000), Kid (Cytoskeleton Inc., IB: 1/1000), KNL1 (Novus Biologicals, IF; 1/500), Mad2 (Novus Biologicals, IF; 1/500), Nde1 (Protein Tech Group, IF; 1/500), Ska1 (Abcam, IF; 1/500), Spindly (Bethyl Laboratories, IF; 1/500, IB; 1/2000), Zwint-1 (Bethyl Laboratories, IF; 1/500), Zw10 (Cosmo Bio, IF; 1/200, IB; 1/2000), pericentrin (Abcam, IF; 1/500).

Human pulmonary artery endothelial cells (HPAEC) and EGM-2 growth media kit were obtained from Lonza. Cells were used at passages 5–8, and all stimulations were done in media containing 2% fetal bovine serum. HKSA was purchased from InvivoGen; antibodies to ICAM1, VCAM1, GFP, CLIP-170, TLR4, and Myc were obtained from Santa Cruz Biotechnology; CLASP2 antibody was received from Millipore Sigma; β-actin and α- and β-tubulin antibodies were from Sigma; phospho-NFκB and SOCS3 antibodies were from Cell Signaling; EB1 antibody was obtained from BD Transduction Laboratories. Texas Red phalloidin and Alexa Flour 488 conjugated secondary antibodies were purchased from Molecular Probes. All other biochemical reagents were obtained from Sigma unless otherwise specified.