Rabbit anti claudin 5 pab

The procedure for tissue preparation and staining was as previously reported^{50 (link)}. Briefly, fixed specimens were embedded in optimal cutting temperature compound (Sakura Finetek, Tokyo, Japan) and sectioned at 20 or 50 μm. For immunohistochemistry, the following antibodies were used: Armenian hamster anti-mouse CD31 mAb (MAB1398Z; Merck Millipore, Darmstadt, Germany; dilution 1/200), rabbit anti–claudin-5 pAb (polyclonal antibody; Abcam, Cambridge, MA, USA; dilution 1/200), rabbit anti-neural/glial antigen-2 (NG2) pAb (AB5320, Sigma–Aldrich, St. Louis, Missouri, USA; dilution 1/250), rabbit anti-glial fibrillary acidic protein (GFAP) pAb (HPA056030; Sigma–Aldrich; dilution 1/1000), Alexa Fluor 546–conjugated goat anti-rabbit IgG pAb (A-11010; Thermo Fisher Scientific; dilution 1/200), and Alexa Fluor 647–conjugated goat anti-Armenian hamster IgG pAb (127-605-160; Jackson ImmunoResearch Laboratories, West Grove, PA, USA; dilution 1/400). The sections were visualized using a Leica TSC SP5 confocal microscope and processed with Leica Application Suite (Leica Microsystems, Wetzlar, Germany) and Adobe Photoshop CC software (Adobe Systems, Mountain View, CA, USA).

Rabbit anti-claudin-1 polyclonal antibody (pAb), mouse anti-claudin-2 monoclonal antibody (mAb), rabbit anti-claudin-3 pAb, mouse anti-claudin-4 mAb, mouse anti-claudin-7 mAb, mouse anti-occludin mAb, rabbit anti-tricellulin pAb, rabbit anti-ZO-1 pAb, rabbit anti-ZO-2 pAb, and rabbit anti-JAM-A pAb were purchased from Invitrogen (Carlsbad, CA, USA). Rabbit anti-claudin-5 pAb and mouse anti-β-actin mAb were purchased from Abcam (Cambridge, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Rabbit anti-ATR pAb, anti-ATR pS428 pAb, mouse anti-Chk1 mAb, and rabbit anti-Chk1 pS345 pAb were purchased from Cell Signaling Technology (Beverly, MA, USA). Goat anti-Rabbit IgG peroxidase-conjugated antibody and goat anti-mouse IgG peroxidase-conjugated antibody were purchased from Millipore (Bedford, MA, USA). Alexa Fluor 488 goat anti-rabbit IgG was purchased from Molecular Probes (Eugene, OR, USA).