Invivoplus rat igg2b isotype control

T-cell depletion was performed by intraperitoneal monoclonal antibody (mAb) injection. Mice were injected with 100 μg of InVivoPlus anti-mouse CD4 (GK1.5, BioXCell) [47 (link)] and/or anti-mouse CD8a mAb (53–6.7, BioXCell) [48 (link)] intraperitoneally at 3 and 1 day prior to LASV infection. InVivoPlus rat IgG2b isotype control and/or InVivoPlus rat IgG2a isotype control (BioXCell) were used as isotype controls. T-cell depleted mice were inoculated with 10³ PFU of LASV in 100 μl of PBS.

Antibodies used included rabbit anti-human PD-L1 (#13684, Cell Signaling Technology, Beverly, MA, USA; 1:1000 for Western blot), anti-Cleaved Caspase-3 (#9664, Cell Signaling Technology; 1:250 for IHC), Rabbit anti-human AhR (#83200, Cell Signaling Technology; 1:50 for ChIP), goat anti-mouse PD-L1 (#AF1019, R&D, Minneapolis, MN, USA; 10 µg/mL for IHC), anti-Ki67 (#ab15580, Abcam, Cambridge, MA, USA; 1:500 for IHC), TTF1 (#ab76013, Abcam, Cambridge, MA, USA; 1:200 for IHC), anti-β-Actin (#A1978, Sigma, St. Louis, MO, USA; 1:5000 for WB), APC anti-mouse CD3 (#100235, Biolegend, San Diego, CA, USA; 1:20 for flow cytometry), PE/Cy7 anti-mouse CD8a (#100721, Biolegend; 1:20 for flow cytometry), APC/Cy7 anti-mouse CD4 (#100413, Biolegend; 1:20 for flow cytometry), PE anti-mouse CD45 (#103105, Biolegend; 1:20 for flow cytometry), PE/Cy7 anti-mouse PD-L1 (#124313, Biolegend; 1:20 for flow cytometry), PE anti-mouse PD-1 (#135206, Biolegend; 1:20 for flow cytometry), FITC anti-mouse B220 (#103205, Biolegend, San Diego, CA, USA; 1:20 for flow cytometry), In Vivo Plus anti-mouse PD-L1 (#BP0101), In Vivo Plus Rat IgG 2b Isotype Control (Clone: LTF-2, #BP0090) were purchased from BioXcell (West Lebanon, NH, USA). Benzo(a)pyrene (#B1760) and Alpha-Naphthoflavone (ANF; # N5757) were purchased from Sigma.

MI-2 (Malt1 inhibitor, Selleckchem.com) was dissolved in DMSO (Sigma-Aldrich, St Louis, MA, USA), aliquoted and stored at -20°C. Albumin from chicken egg white (OVA, Sigma-Aldrich, St Louis, MA, USA) and Poly (I:C) (HMW) (In vivoGen, San Diego, CA, USA) were aliquoted and stored at -20°C. Diphtheria Toxin (DT) from Corynebacterium diphtheriae (Sigma-Aldrich, St Louis, MA, USA) was aliquoted and stored at -80°C. Anti-mouse PD-L1 antibody InVivoPlus (B7-H1) (BioXcell, Lebanon, NH, USA) and InVivoPlus rat IgG2b isotype control, (BioXcell, Lebanon, NH, USA) were stored at 4°C.