Anti phospho pyk2 tyr402

Whole cell lysates were prepared in M-PER buffer (Thermo, Waltham, MA), resolved by SDS-PAGE and blotted with indicated antibodies. The following antibodies were used in this study: Anti-HA, anti-FLAG, anti-phospho-β-catenin (Ser33/37/Thr41), anti-β-catenin, anti-GSK3α, anti-β-TrCP, anti-α-Tubulin, anti-GSK3β, anti-FAK, anti-phospho-FAK (Tyr397), anti-PYK2 and anti-phospho-PYK2 (Tyr402) (Cell Signaling); anti-FLAG M2 (Sigma); anti-ubiquitin and anti-β-catenin (BD Bioscience, San Jose, CA); anti-GSK3, anti-c-Myc, anti-PYK2, anti-β-TrCP, anti-phospho-GSK3β (Tyr216) and anti-β-Actin (Santa Cruz Biotechnology, Dallas, TX); anti-phospho-GSK3 (Tyr279/Tyr216) (Millipore). anti-GSK3β for IP was purchased from Abcam (Cambridge, MA) and Bethyl Laboratories (Montgomery, TX). SuperSignal West Pico Chemiluminescent Substrate and SuperSignal Western Blot Enhancer (Thermo) were used to enhance western signal when needed. PF-562271 was purchase from MedKoo Biosciences (Chapel Hill, NC) and Selleck Chemicals (Houston, TX). Gelucire is a gift from Gelucire-4414.html">Gattefosse (Paramus, NJ). MG-132 was obtained from Sigma.

Recombinant human VCAM-1/CD106 Protein and Recombinant Human/Rhesus Macaque/Feline CXCL12/SDF1α were from R&D Systems (Minneapolis, MN, USA). Human fibronectin, doxorubicin and FAK inhibitor PF-228 were purchased from Millipore Sigma (Oakville, ON, Canada). The PYK2 inhibitor VS-6063 was purchased from Cayman Chemicals (Ann Arbor, MI, USA). Anti-phospho-PYK2 (Tyr-402) and anti-PYK2 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The fluorescein isothiocyanate (FITC)-conjugated Annexin V, Phycoerythrin (PE)-conjugated anti-human α4 integrin, Alexa 647-conjugated anti-human α5 integrin antibodies, FITC-conjugated anti-human CD3 and PE-conjugated anti-mouse IgG were purchased from BD Biosciences (San Jose, CA, USA). Anti-human β1 integrin (4B4) was purchased from Beckman Coulter Life Sciences (Mississauga, ON, Canada).