Mouse anti rac1

We used the following primary antibodies: mouse-anti-RhoA (26C4, Santa Cruz, Heidelberg, Germany, sc-418), mouse-anti-Rac1 (BD Transd. Laboratories, Heidelberg, Germany, 610650), mouse-anti-p190RhoGAP (p190A) (BD Transd. Laboratories, 610149), mouse-anti-p190B (BD Transd. Laboratories, 611612), mouse-anti-RGS3 (CC-Q7, Santa Cruz, sc-100762), mouse-anti-caveolin-3 (BD Transd. Laboratories, 610421), mouse immunoglobulin G (IgG) (Santa Cruz sc-2025), rabbit-anti-caveolin-3 (H-100, Santa Cruz, sc-28828), mouse-anti-eNOS (BD Transd. Laboratories, 610297), rabbit-anti-phospho-1177Ser-eNOS (Cell Signaling Technology, Frankfurt, Germany, #9571), rabbit-anti-Nitrotyrosine (Millipore, Darmstadt, Germany, 06-284). The corresponding horseradish peroxidase-conjugated secondary antibodies were from Sigma-Aldrich (Munich, Germany, A-9044, A-9169). In this study, the following reagents and inhibitors were used: carbamoylcholine chloride (carbachol, Sigma-Aldrich, Munich, Germany, C4382), 5-bromo-2′-deoxyuridine (5-BrdU) (Sigma-Aldrich), methyl-β-cyclodextrin (MβCD, Sigma-Aldrich, M7439), (2S)-2-amino-5-(1-aminoethylideneamino)pentanoic acid; dihydrochloride (L-NIO dihydrochloride, Santa Cruz, sc-361229), Nω-nitro-L-arginin-methylester-hydrochloride (L-NAME, Sigma-Aldrich, 72760).

All SDS-PAGE and Western blotting procedures were carried out according to standard protocols. The following primary antibodies were used: mouse anti-UCH-L1 (31A3, a gift from Professor Ian Day, University of Bristol, and Prof. Rod Thompson, 1:10,000), rabbit anti-GluA1 (Millipore, 1:1000), rabbit anti-RhoGDI (Abcam, 1:5000), mouse anti-H-ras (Santa Cruz Biotechnology, 1:1000), mouse anti-Rac1 (Santa Cruz Biotechnology, 1:1000), rabbit anti-HA (Abcam, 1:5000), goat anti-Lamin B (Santa Cruz Biotechnology, 1:2000), and rabbit anti-PSD95 (Abcam, 1:1000). All blots were developed using Li-Cor Odyssey imaging software, except for anti-Lamin B, which was developed on film using HRP-conjugated anti-goat (Sigma, 1:10,000). For Li-Cor Odyssey Western blot imaging, the IRDye secondary antibodies (all from Li-Cor) used were 800CW donkey anti-mouse, 800CW donkey anti-rabbit, 680RD donkey anti-mouse, and 680RD donkey anti-rabbit, all 1:10,000. Western blot signal quantification was performed using Li-Cor Odyssey imaging software.