Western blotting materials

Endothelial cell growth medium (EGM), extracellular matrix gel (ECM), TRIzol, RIPA buffer, collagenase A, and AATF antibody were procured from Sigma-Aldrich (St. Louis, USA). Dulbecco’s modified eagle’s medium (DMEM), giemsa stain, hematoxylin, 8μm-transwell inserts, and 24- well plates were procured from HiMedia laboratories (India). Lipofectamine 3000, penicillin/streptomycin, glutamine, FBS were from Invitrogen (USA); WST-1 cell proliferation assay kit from Takara Bio Inc. (Shinga, Japan); antibodies to IgG, PEDF, pAkt, pErk1/2, pFAK, Akt, Erk, and control and AATF shRNA from Santa Cruz Biotechnology, Inc. (CA, USA); antibodies to β-actin from Cell Signaling Technologies (USA); verso cDNA synthesis kit, and DyNamo Colorflash SYBR green kit from Thermo Fisher Scientific (USA); western bright ECL HRP substrate from Advansta (USA); western blotting materials were from BioRad; primers from Integrated DNA Technologies (IDT) (Iowa, USA), and all other reagents were obtained from Thermo Fisher Scientific or Sigma.

Cell culture media and reagents were from Lonza (Basel, Switzerland). Western blotting materials were obtained from Bio-Rad (Hercules, California). Akt inhibitor VIII and PKCζ pseudosubstrate inhibitor myristoylated were from Calbiochem, (Darmstadt, Germany). PI3K inhibitor (LY294002), rapamycin and torin-1, as well as antibodies against phospho-STAT6 (Tyr641), phospho-Akt (Ser 473), phospho-PKCζ/λ (Thr410/403), phospho-4E-BP1 (Ser65), phospho-GSK3β (Ser 9), phospho-FoxO3a (Thr32), their respective total protein antibodies, and α-tubulin were purchased at Cell Signaling Technologies (Danvers, Massachusetts). Antibodies against anti-β actin, anti-PKCζ H1 and anti-EEA1 were from Santa Cruz Biotechnology (Dallas, Texas), and anti-STAT6, Alexa Fluor 488 anti-mouse IgG, and Alexa Fluor 633 anti-rabbit IgG were from Thermo Fisher Scientific (Waltham, Massachusetts). All other reagents were of analytical grade, purchased from Sigma-Aldrich (St. Louis, Missouri) unless otherwise specified.