Mouse anti occludin mab

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).

n-Dodecyl-β-D-maltoside (DDM) was purchased from Dojindo Laboratories (Kumamoto, Japan). FD-4 was purchased from Sigma-Aldrich (St. Louis, MO, USA). Monoclonal anti-6 histidine antibody conjugated europium³⁺ cryptate (Eu(K)-anti-His Ab) and monoclonal anti-GST antibody labelled with XL665 (XL665-anti-GST Ab) were purchased from CIS Bio International (Gif-sur-Yvette, France). Rabbit anti-claudin-1 polyclonal antibody (pAb), rabbit anti-claudin-3 pAb, mouse anti-claudin-4 monoclonal Ab (mAb), mouse anti-occludin mAb and rabbit anti-ZO-1 pAb were purchased from Invitrogen (Carlsbad, CA, USA). Mouse anti-β-actin mAb was purchased from Sigma-Aldrich. 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 and Alexa Fluor 488 goat anti-mouse IgG were purchased from Molecular Probes (Eugene, OR, USA).
The human colorectal adenocarcinoma cell line Caco-2 (HTB-37) was obtained from the American Type Culture Collection (Rockville, MD, USA). Caco-2 cells were cultured in Eagle’s minimum essential medium (Nissui, Japan) supplemented with 10% foetal bovine serum under 5% CO₂ at 37 °C. The passage number of the cells used for the experiments was between 10 and 20.

RhoA inhibitor (CT04) was obtained from Cytoskeleton Inc. (Denver, CO, USA), and ROCK inhibitor (Y27632), MLC inhibitor (blebbistatin), MLCK inhibitor (ML7), and PKC inhibitor (Gö 6983) were purchased from Sigma-Aldrich. SlowFade Gold antifade reagent with 4′,6-diamidino-2-phenylindole (DAPI) was obtained from Molecular Probes (Bedford, MA, USA). Specific rabbit polyclonal antibodies against pMLC (Ser19), MYPT, and pMYPT (at Thr853) and rabbit MAbs against MLC2, pan-pPKC, and ROCK were purchased from Cell Signaling (Beverly, MA, USA). Rabbit anti-MLCK MAb was obtained from Abcam (Cambridge, MA, USA). Mouse antioccludin MAb was purchased from Invitrogen (Carlsbad, CA, USA); rabbit anti-claudin and anti-JAM polyclonal antibodies and rabbit anti-ZO-1 polyclonal antibody were obtained from Life Technologies (Eugene, OR, USA). Mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (0411) MAb was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Secondary antibodies included horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (Cell Signaling), HRP-conjugated goat anti-mouse IgG (Santa Cruz Biotechnology), and AF488-conjugated donkey anti-rabbit IgG and AF488-conjugated goat anti-mouse IgG (Life Technologies).