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2 protocols using anti pd 1 antibody pembrolizumab

1

Immune Checkpoint Inhibitor Screening Assays

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Caffeic acid (#CFN99190), luteolin (#CFN98784), luteolin 7-O-glucoside (#CFN98565), cosmosiin (#CFN98981), homoplantaginin (#CFN90344), apigenin (#CFN98843), and hispidulin (#CFN99491) were purchased from ChemFaces (Wuhan, China). PD-1/PD-L1 inhibitor screening assay kit (#72005), CTLA-4 (cytotoxic T-lymphocyte-associated protein 4)/CD80 inhibitor screening assay kit (#72009), and PD-L1-blocking antibody (#71213) were purchased from BPS Bioscience (San Diego, CA, USA). Small molecule PD-L1 inhibitor C1 (BMS-1, # T3655) was purchased from TargetMol (Wellesley Hills, MA, USA). Anti-PD-1 antibody (Pembrolizumab, #A2005) and anti-CTLA-4 antibody (#A2001) was purchased from Selleck Chemicals (Houston, TX, USA).
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2

Culturing and Characterizing NK Cell Lines

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Natural-killer cell line NK-92 (Cat# CRL-2407) was available from American Type Culture Collection (Manassas, VA, USA). SNK-6 was kindly provided by Professor Norio Shimizu and Yu Zhang of Chiba University. Cell lines were authenticated using Short Tandem Repeat (STR) analysis (Genetic Testing Biotechnology, STR Profile Reports in Supplemental Data). Recent mycoplasma testing has been performed with Lonza LT07-705 MycoAlert™ PLUS Mycoplasma Detection Kit. NK-92 cells were cultured in α-MEM medium supplemented with 10% FBS, 10% HBS and recombinant human IL-2 (20ng/ml). SNK-6 cells were cultured in RPMI-1640 medium supplemented with 10% FBS, 10% HBS and recombinant human IL-2 (80ng/ml). Asparaginase (Cat# A3809), 13C-glutamine (Cat# 184161-19-1), and glutamine (Cat# 56-85-9) were obtained from Sigma-Aldrich (St.Louis, MO, USA). Anti-PD-1 antibody pembrolizumab (Cat# A2005) was from Selleck Chemicals (Houston, TX, USA).
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