Claudin 5 and zo 1

Antibodies were obtained from the following commercial sources: Matrix metalloproteinase- (MMP-) 9 and histone deacetylase- (HDAC-) 4 (Cell Signaling Technology), claudin-5 and ZO-1 (Invitrogen), occludin, the nicotinamide adenine dinucleotide phosphate oxidase- (NOX-) 4, and GAPDH (Proteintech), and rhodamine phalloidin was obtained from Cytoskeleton. GKT137831 and Tasquinimod (Taq) was from selleck. Lactate dehydrogenase (LDH) activity assay kit was bought from Beyotime Institute of Biotechnology. Cell counting kit 8 (CCK8) was from Dojindo. 2,3,5-Triphenyltetrazolium chloride (TTC), 2′,7′-dichlorofluorescin diacetate (DCFH-DA), diphenyleneiodonium (DPI), fluorescein isothiocyanate- (FITC-) labeled dextran, and other chemical reagents were from Sigma-Aldrich.

Immunohistochemistry was performed on 10-μm frozen sections of cold phosphate buffer saline (PBS)-perfused tissues as described previously [33 (link)]. Antibodies reactive for the following antigens were used in this study: rat monoclonals reactive to CD31 (MEC13.3), β4 integrin (346-11A), MHC class II (M5/114.15.2), CD45, Mac-1 (M1/70), all from BD Pharmingen (La Jolla, CA); mouse monoclonal α-SMA-Cy3 conjugate (1A4) from Sigma (St. Louis, MO); rabbit polyclonals reactive to claudin-5 and ZO-1 from Invitrogen (Carlsbad, CA); and fibrinogen from Millipore (Temecula, CA). Secondary antibodies used included Cy3-conjugated anti-rat and anti-rabbit from Jackson Immunoresearch (West Grove, PA) and anti-rat Alexa Fluor 488 and anti-rabbit Alexa Fluor 568 from Invitrogen (Carlsbad, CA).