Anti claudin 3

Anti-ZO-1, anti-occludin, and anti-claudin-3 antibodies were purchased from Invitrogen (Carlsbad, CA). Anti E-Cadherin and anti β-catenin antibodies were purchased from BD Biosciences (Billerica, MA). Horseradish peroxidase (HRP)-conjugated anti-mouse IgG, HRP-conjugated anti-rabbit IgG and anti-β-actin antibodies were obtained from Sigma Aldrich (St. Louis, MO). AlexaFlour-488-conjugated anti-mouse IgG and Cy3-conjugated anti-rabbit IgG were purchased from Molecular Probes (Eugene, OR).

Cultured cells were rinsed twice with ice-cold HBSS, lysed in protein loading buffer (50 mM Tris, pH 6.8, 100 mM dithiothreitol, 2% SDS, 0.1% bromophenol blue, 10% glycerol), and then sonicated. Mouse colonic epithelial cells were collected by scraping the tissue from the colon of the mouse, including the proximal and distal regions.^{51 (link), 52 (link)} The cells were sonicated in lysis buffer (10 mM Tris, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, pH 8.0, 1% Triton X-100) with 0.2 mM sodium ortho-vanadate, and protease inhibitor cocktail. The protein concentration was measured using the BioRad Reagent (BioRad, Hercules, CA, USA). Equal amounts of protein were separated by SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and immunoblotted with primary antibodies. The following antibodies were used: anti-claudin-2, anti-claudin-3, anti-claudin-7 (Invitrogen, Carlsbad, CA, USA), anti-p-STAT3, anti-STAT3, anti-p-IκBα, anti-IκBα, anti-p-SAPK/JNK, anti-SAPK/JNK (Cell Signal, Beverly, MA, USA), anti-Villin and anti-VDR (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) or anti-β-actin (Sigma-Aldrich, Milwaukee, WI, USA) antibodies and were visualized by ECL. Membranes that were probed with more than one antibody were stripped before re-probing.

Sodium butyrate, potato starch (PS; Sigma-Aldrich, St. Louis, MO). All primers for quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) were synthesized by Integrated DNA Technologies (Coralville, IA). Antibodies were purchased from the following sources: anti–Na⁺/H⁺ exchanger isoform 3 (NHE₃), anti–zona occludin-1 (ZO-1), anti–sodium-coupled monocarboxylate transporter 1 (SLC5A8), and anti–frizzled class receptor 7 (FZD7) from Abcam (Cambridge, MA); anti-occludin from Hycult Biotechnologies (Plymouth Meeting, PA); anti-claudin-3, Alexa Fluor 488, and 568 IgG from Invitrogen (Camarillo, CA).

Anti-ZO-1, anti-occludin, and anti-claudin-3 antibodies were purchased from Invitrogen (Carlsbad, CA). Anti E-Cadherin and anti β-catenin antibodies were purchased from BD Biosciences (Billerica, MA). Horseradish peroxidase (HRP)-conjugated anti-mouse IgG, HRP-conjugated anti-rabbit IgG and anti-β-actin antibodies were obtained from Sigma Aldrich (St. Louis, MO). AlexaFlour-488-conjugated anti-mouse IgG and Cy3-conjugated anti-rabbit IgG were purchased from Molecular Probes (Eugene, OR).

Anti-ZO-1 (rabbit polyclonal), anti-occludin (mouse monoclonal), anti-claudin-2 (mouse monoclonal) and anti-claudin-3 (rabbit polyclonal) antibodies were purchased from Invitrogen (Carlsbad, CA). Rabbit polyclonal anti-toll like receptor-4 (TLR4) antibody was from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti E-Cadherin (mouse monoclonal) and anti β-catenin (rabbit polyclonal) antibodies were purchased from BD Biosciences (Billerica, MA). Horseradish peroxidase (HRP)-conjugated anti-mouse IgG, HRP-conjugated anti-rabbit IgG and anti-β-actin (mouse monoclonal) antibodies were obtained from Sigma Aldrich. AlexaFlour-488-conjugated anti-mouse IgG and Cy3-conjugated anti-rabbit IgG were purchased from Molecular Probes (Eugene, OR).