Rat anti cd31 clone mec13.3

Recombinant human VEGF₁₆₅ (Ala27-Arg191, no. 293-VE), human Sema3C (Gly21-Ser738, no. 5570-S3), and mouse Sema3C (Gln24-Ser741, no. 1728-S3) were from R&D Systems. Recombinant human IgG-Fc (009-000-008) was purchased from Jackson ImmunoResearch. Antibodies: rat anti-Sema3C (clone 238835) and goat anti-Nrp-1 (AF566) are from R&D Systems; rabbit anti-pY397FAK, rabbit anti-pT180/pY182-p38MAPK (Invitrogen); rabbit anti-FAK (C-20), and goat anti-Nrp-1 (C-19) are from Santa Cruz; mouse anti-p38MAPK (L53F8), rabbit anti-AKT, rabbit anti-pS473AKT, and rabbit anti-cleaved Caspase-3 (Asp175) are from Cell Signaling; pTyr118 paxillin (NEB), rat anti-CD34 (NCL-END; Novocastra); rat anti-CD31 (clone Mec13.3) and rat anti-VE-cadherin are from BD Biosciences; mouse anti-vinculin (V9131) and anti-actin smooth muscle-Cy3 are from Sigma; rabbit anti-GFAP (DAKO), rabbit anti-NG2 (Millipore), rabbit anti-collagen IV (Cosmo Bio).

For immunofluorescence analysis, the following primary antibodies were used: rabbit anti‐ZO‐1 (1:100 dilution, Thermo Fisher, #61‐7300), rabbit anti‐claudin‐5 (1:400, Abcam, #ab53765), rabbit anti‐GFAP (1:500, DAKO, #Z0334), rat anti‐CD31 (clone Mec13.3, BD Biosciences), Pals1 (1:100, Merck, #7‐708), rat anti‐Crb3 (14F9, Abcam, ab180835), mouse anti‐E‐cadherin (1:200, BD Bioscience, #610181), and rabbit anti‐cleaved caspase‐3 (Asp175, 1:100 Cell Signaling, #9661). Secondary antibodies were 3 mg/ml Alexa 546‐conjugated goat anti‐rabbit IgG and 3 mg/ml Alexa 488‐conjugated goat anti‐rat IgG (all from Invitrogen). For Western blotting, rabbit anti‐MPDZ (1:500, Thermo Scientific, 42‐2700) and mouse anti‐beta actin (1:2,500, Sigma‐Aldrich #A5441) were used.