High glucose dulbecco s modified eagle s medium dmem

The present study was approved by the Medical Ethics Committee of the West China Hospital of Stomatology of Sichuan University (Chengdu, China). Specimens of OSCC and normal oral mucosal tissue were obtained from 5 patients (2 males, 3 females; mean age, 61.6 years) from November 2015 to December 2015 at the Department of Head and Neck Oncology, West China Hospital of Stomatology, and all patients provided informed consent prior to inclusion in the current study. The isolation and characterization of primary CAFs and NFs was performed according to a previously established protocol (8 (link)). CAFs and NFs were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM; GE Healthcare, Chicago, IL, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc., Waltham, MA, USA), penicillin (100 U/ml) and streptomycin (100 µg/ml; GE Healthcare) at 37°C in a humidified incubator with 5% CO₂. HLECs were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in endothelial cell medium (ECM; ScienCell Research Laboratories) supplemented with endothelial cell growth supplement (ScienCell Research Laboratories), 10% FBS, penicillin (100 U/ml) and streptomycin (100 µg/ml) at 37°C in a 5% CO₂ humidified atmosphere. Cell culture duration is described in each specific experiment as follows.

Methyl-β-cyclodextr in (MβCD) and anti-neutrophil myeloperoxidase [MPO; mouse anti-goat polyclonal IgG (H+L)] antibodies were obtained from Santa Cruz Biotechnology, Inc. (1:100, cat. no. sc-16129, Dallas, TX, USA), along with lipid-raft disruptor filipin. Alexa Fluor 488-cholera toxin subunit B (CTXB) and Alexa Fluor 594-labeled chicken anti-Goat IgG (H+L) secondary antibody were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Anti-flotillin-1 antibody was purchased from BD Biosciences (1:1000, cat no. 61802; Franklin Lakes, NJ, USA). Polyclonal anti-ERp29 rabbit anti-mouse antibody was obtained from Abcam (1:3000, ab11420; Cambridge, MA, USA). Optiprep was obtained from Axis-Shield, Inc. (Norton, MA, USA). High glucose Dulbecco's modified Eagle's medium (DMEM) was purchased from GE Healthcare Life Sciences (Logan, UT, USA). An Enhanced Chemiluminescence (ECL) kit was obtained from PerkinElmer Inc. (Waltham, MA, USA). Human LDL was purchased from ProSpec-Tany TechnoGene, Ltd. (Ness Ziona, Israel).