Egmtm 2 mv microvascular endothelial cell growth medium 2 bulletkit

Hec50 cells were kindly provided by Dr. Erlio Gurpide (New York, NY, USA) [18 (link)] and grown in high-glucose Dulbecco’s Modified Eagle Medium (DMEM; Gibco Corporation, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum. KLE cells were purchased from ATCC and grown in RPMI-1640 medium supplemented with 10% fetal bovine serum. Human umbilical vascular endothelial cells (HUVEC) were purchased from ATCC and grown in EGMTM -2 MV Microvascular Endothelial Cell Growth Medium (2 BulletKit, Lonza, Alpharetta, GA, USA). To ensure rigor and reproducibility, the identity of all cell lines was confirmed using the CODIS genotyping test (Cat. No. CL1003, Bio-Synthesis, Lewisville, TX, USA).

The following reagents and antibodies were used in this study: p21 Waf1/Cip1 (12D1) rabbit, p16 INK4A (D7C1M) rabbit, p53 (1C12) mouse (Cell Signaling Technology, Frankfurt, Germany); anti-phospho-IRE1 (S724) rabbit monoclonal antibody (Boster, Germany); human XBP1 antibody (R & D System, Wiesbaden, Germany); anti ATF6 alpha (Rockland, Germany); alexa fluor FITC goat anti-rabbit IgG, alexa fluor TRITC goat anti-rabbit IgG, alexa fluor TRITC goat anti-mouse IgG and alexa fluor FITC goat anti-mouse IgG (Invitrogen, Karlsruhe, Germany). Human coronary artery endothelial cells were purchased from Lonza Germany (Catalog #: CC-2585); EGMTM -2 MV microvascular endothelial cell growth medium-2 bulletkit (Lonza, Köln, Germany); D-(+)-Glucose (Sigma, Darmstadt, Germany); human oxidized low density lipoprotein (oxLDL) (Thermo Fisher Scientific, Dreieich, Germany); vectashield mounting medium with DAPI (Vector Laboratories, Newark, CA, USA); streptozotocin (Enzo Life Sciences, Lörrach, Germany); “high-fat diet” (HFD) experimental food (Western Type Diet, 43% carbohydrates, 15% proteins and 42%, Ssniff, Soest, Germany); accu-chek test strips, accu-check glucometer; albumin fraction V (Carl Roth, Karlsruhe, Germany); rompun 2% (Bayer, Leverkusen, Germany); ketamine 10% (beta-pharm, Augsburg, Germany); chemiluminescent western blot reagents (Merk Millipore, Darmstadt, Germany).