Phospho vasp ser239

Linaclotide was provided by Ironwood Pharmaceuticals Inc (Cambridge, MA). EZ‐Link Sulfo‐NHS‐SS‐Biotin and streptavidin‐agarose resins were obtained from Thermo Scientific (Rockford, IL). 8‐Bromoguanosine 3′,5′‐cyclic monophosphate sodium salt monohydrate (8Br‐cGMP), bumetanide, GlyH‐101, and NKH477 were obtained from Sigma‐Aldrich (St. Louis, MO). The PKA inhibitor H7 and PKG inhibitor H8 dihydrochloride were purchased from AbCam (Cambridge, MA). The mouse monoclonal CFTR‐450 antibody raised against full‐length human CFTR was obtained from Cystic Fibrosis Foundation Therapeutics, University of North Carolina‐Chapel Hill. The affinity‐purified polyclonal antibody AME4991 raised against rat CFTR was generated by Dr. Ameen (Ameen et al. 2000; Golin‐Bisello et al. 2005; Jakab et al. 2011). Phospho‐VASP (Ser239) and antimouse IgG, HRP‐linked antibodies were obtained from Cell Signaling Technology Inc. (Danvers, MA). The cGKII (PRKG2) rabbit polyclonal antibody was obtained from Proteintech Group Inc. (Chicago, IL) and β‐actin monoclonal antibody from Sigma‐Aldrich (St. Louis, MO). All other drugs, reagents, and chemicals were obtained from Sigma‐Aldrich unless otherwise stated.

Antibodies for immunoblot analyses included the following: BiP, NPY, phospho-VASP^Ser239, phospho-VASP^Ser157, β-actin, β-tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and villin-1 (Cell Signaling Technology, Danvers, MA, USA); Gucy2c (MS20)^{8 (link)} and prouroguanylin (6910, M Goy, University of North Carolina).^{11 (link)} Horseradish peroxidase-conjugated secondary antibodies were from Santa Cruz Biotechnology (Dallas, TX, USA) or Jackson ImmunoResearch Laboratories (West Grove, PA, USA), and SuperSignal West Dura Chemiluminescent Substrate and Femto Chemiluminescent Substrate were from Thermo. Band intensities were quantified by densitometry and normalized to β-actin, β-tubulin, GAPDH or villin-1.