Mouse anti actin

Western blot analyses were performed as previously described (Wang et al., 2011 (link)). Animals were anesthetized and sacrificed by cervical dislocation. The brain tissue was immediately harvested and stored at −80°C. The tissue dissected from mouse brain was homogenized in RIPA buffer. After centrifuging at 12,000 rpm for 5 min, supernatant was collected as the total cell lysate. Protein concentration was measured using the BCA Protein Assay Kit from Thermo Fisher Scientific. Supernatant was separated on 4%–15% polyacrylamide Tris-HCl gradient gels (BioRad, Hercules, CA, USA) and transferred to PVDF membranes. The integrated optic density of the immunoreactive band was quantified with NIH ImageJ software. The primary antibodies used were: rabbit polyclonal superoxide dismutase 2 (SOD2; 1:2000; EMD Millipore), rabbit polyclonal glutathione peroxidase 1 (GPx1; 1:1000; Abcam, Cambridge, MA, USA), goat polyclonal NAD(P)H:Quinone Oxidoreductase 1 (NQO1; NQP1; 1:1000; Abacam), rabbit polyclonal heme oxygenase 1 (HO1; 1:1000; Enzo Life Sciences, Farmingdale, NY, USA) and mouse anti-actin (1:5000, EMD Millipore).

Dulbecco's modified Eagle's medium (DMEM), phosphate-buffered saline (PBS), Hank's balanced salt solution (HBSS), fetal bovine serum (FBS), normal goat serum (NGS), and antibiotic-antimycotic were purchased from Gibco (Grand Island, NY, USA). Trimethyltin chloride, 2,7-dichlorofluorescin diacetate (DCFH-DA), 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), Tween-20, Hoechst 33258, SB203580, SP600125, apocynin, and BAY11-7082 were from Sigma-Aldrich (St. Louis, MO, USA). Rabbit antibodies against p-JNK, p-p38, p-ERK, p-IκBα, p38, and IκBα were purchased from Cell Signaling Technologies, Inc. (Beverly, MA, USA). Rabbit anti-JNK, ERK, and NF-κB p65 antibodies were from Santa Cruz Biotechnology (Santa, CA, USA). Mouse anti-inducible nitric oxide synthase (iNOS) was from BD Biosciences (Franklin Lakes, NJ, USA). Mouse anti-actin was purchased from EMD Millipore (Billerica, MA, USA). Rat anti-CD11b was from AbD Serotec (Oxford, UK). The goat anti-mouse, -rabbit, and -rat IgG (HRP-conjugated) secondary antibodies were from Enzo Life Science (Farmingdale, NY, USA). All other chemicals were purchased from Sigma-Aldrich.

All materials were purchased from Sigma (St Louis, MO) unless stated otherwise. Tie2-Cre plasmid was a gift from Dr. Masashi Yanagisawa, UT Southwestern (Tie2 is endothelium-specific [20 (link)]). Sheep anti-mouse GBM antibody was a gift from Dr. Hui Lan (Chinese University of Hong Kong). Goat anti-hSTC1 antibodies and rabbit anti-AQP1 antibodies were purchased from Santa-Cruz Biotechnology, Inc. (Santa Cruz, CA). Rabbit anti-CD3 antibodies, mouse anti-actin and mouse anti-GAPDH were purchased from EMD Millipore (Billerica, MA). Rat anti-F4/80 antibodies were purchased from AbD Serotec (Raleigh, NC). Rabbit anti-sheep IgG and rabbit anti-mouse IgG were purchased from Bethyl Laboratory (Montgomery, TX). Rabbit anti-mouse C3 was purchased from Gene Tex (Irvine, CA). Anti-mouse-IgG,-IgG₁,-IgG_2a,-IgG_2b and-IgG₃ antibodies were purchased from Southern Biotechnology Associates (Birmingham, AL). ECL plus reagent was purchased from Fisher Scientific (Pittsburgh, PA).

All materials were purchased from Sigma Aldrich Inc. (St Louis, MO) unless stated otherwise. Recombinant human STC1 (rSTC1) was purchased from MyBioSource (San Diego, CA). MnTBAP, rat anti-F4/80 (macrophage marker), rabbit anti-CD3 (T-cell marker), mouse anti-actin and mouse anti-GAPDH were purchased from EMD Millipore (Billerica, MA). SCH79797 was purchased from Fisher Scientific (Houston, TX). Goat anti-hSTC1 antibodies and Goat anti-thrombin antibodies were purchased from Santa Cruz (Santa Cruz, CA). Rabbit anti-ERK and rabbit anti-p-ERK were purchased from Cell Signaling (Danvers, MA).