Dichlorofluorescin diacetate dcf da

C57BL/6 WT, Pmel trangenic and Rag1^−/− mice were purchased from the Jackson Laboratory. Dr. Hans Schreiber (University of Chicago) provided the B16F10, B16-SIY cell lines, SIY peptides and 2C transgenic mice. CD73^−/− mice have been described previously (22 (link)). ID8-OVA cells were generated as described previously (23 (link)). All the cell lines were routinely tested for mycoplasma infections by culture and DNA stain, and maintained in complete medium composed of RPMI 1640 with 5% FBS. All animal experiments were approved by institutional animal use committees of the Northwestern University. Eflornithine (α-difluoromethylornithine or DFMO) was purchased from AK Scientific, Inc. Dichlorofluorescin diacetate (DCFDA), and 5-fluorouracil (5-FU) were purchased from Sigma-Aldrich. CD73 selective inhibitors APCP and CD39 selective inhibitors ARL67156 were from Tocris Bioscience. Arginase 1/ARG1 Fluorescein-conjugated antibodies was purchased from R&D Systems. All the mAbs for flow cytometry were purchased from eBioscience and BioLegend. The D^b/gp100 tetramers were provided by the National Institutes of Health Tetramer Core Facility (Atlanta, GA). Depleting mAb clone 53.6.7 (anti-CD8α) and RB6-8C5 (anti-Gr1) antibodies were purchased from Bio X Cell. N^ω-hydroxy-nor-Arginine (Nor-NOHA) and arginase I activity kit were purchased from Cayman Chemical Company.

BaP (Sigma, MO, USA), gefitinib (Santa Cruz Biotechnology, CA, USA), and EGCG (≥95%, Sigma, MO, USA) were all dissolved in dimethylsulfoxide (DMSO, MO, USA) to make 39.6 mM, 50 mM, and 50 mM stock solutions, respectively. LHC-9 growth medium, penicillin/streptomycin, and fetal bovine serum (FBS) were purchased from Thermo Scientific (PA, USA). Hyclone (1×) porcine trypsin used for subculture was purchased from GE Healthcare Life Sciences (UT, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Usb Corporation (CA, USA) and trypan blue was from Sigma (MO, USA). Dichlorofluorescin diacetate (DCFDA) was purchased from Sigma (MO, USA) and phosphate buffered saline (1×PBS) was purchased from Gibco Life Technologies (NY, USA). RNeasy^® Blood and Tissue Mini kits were purchased from QIAGEN (MD, USA). One-step real time-polymerase chain reaction (RT-PCR) kits with SYBR^® green were purchased from Bio-Rad (CA, USA). Primers were purchased from Eurofins (Luxembourg) and Western blot antibodies were purchased from Santa Cruz (CA, USA), Abcam (MA, USA), and GE Healthcare (NJ, USA). Radioimmunoprecipitation (RIPA) lysis buffer was from Santa Cruz Biotechnology (CA, USA) and the enhanced chemiluminescence (ECL) kit was purchased from GE Healthcare (NJ, USA).