Anti pd l1 e1l3n

For protein immunoprecipitation (i.p.), the following antibodies were used: mouse monoclonal anti-Ac-lysine (AKL5C1) (sc-32268, Santa Cruz Biotechnology) and rabbit polyclonal anti-PD-L1 (E1L3N) (13684, Cell Signaling).
The following antibodies were used to evaluate protein expression: rabbit polyclonal anti-PD-L1 (E1L3N) (1:500) (13684, Cell Signaling Technology), mouse monoclonal anti-TRAPPC4 (C-7) (1:100) (sc-390551, Santa Cruz Biotechnology) and rabbit polyclonal anti-acetyl-Histone H3 (Lys27) (1:500) (4353, Cell Signaling). (Mouse monoclonal anti-β-actin (1:10,000) (Sigma Aldrich) and rabbit polyclonal anti-Lamin A/C (1:1000) (20 32, Cell Signaling Technology) were used as loading controls. Goat anti-mouse IgG-HRP (1:20,000) (Bethyl Laboratories, A90-116P) and goat anti-rabbit IgG-HRP (1:20,000) (Bethyl Laboratories, A120-101P, Montgomery, TX, USA) were used as secondary antibodies. All primary and secondary antibodies were diluted in a PBS + 0.1% Tween20 solution containing 3% BSA (SERVA, Reno, NV, USA).

Anti-RIPK3 (ab72106), anti-MLKL (ab184718), anti-pMLKL (ab187091), and anti-FOXp3 (ab20034) were purchased from Abcam (Cambridge, UK); anti-PD-L1 (E1L3N) was obtained from Cell Signaling Technology (Danvers, Massachusetts, USA); anti-RIPK1 (610459) was bought from BD Biosciences (San Jose, California, USA); anti-CD8 (M7103) was purchased from Dako (Agilent) (Santa Clara, California, USA); and anti-CD163 (NCL-L-CD163) was purchased from Leica Biosystems (Wetzlar, Germany).

Sub-confluent cells were lysed in Laemmli buffer (LB) and 45 µg of total proteins were subjected to SDS-PAGE and western blotting following standard methods. Protein detection was performed by using the following primary antibodies, diluted as indicated: anti-human MET (3D4, 1:3000, Invitrogen Corp., Camarillo, CA), anti-pMET Tyr^1234/1235 (D26, 1:1000), anti-JAK1 (6G4, 1:1000), anti-pJAK1 Tyr^1022/1023 (1:1000), anti-JAK2 (D2E12, 1:1000), anti-pJAK2 Tyr^1007/1008 (1:1000) anti-STAT1 (1:1000), anti-pSTAT1 Tyr⁷⁰¹ (58D6, 1:1000), anti-PD-L1 (E1L3N, 1:1000), anti-GAPDH (D4C6R, 1:1000) (all from Cell Signaling Technology, Beverly, MA), anti-IFNGR1 (EPR7866, 1:1000, Abcam, Cambridge, UK) and anti-Vinculin (hVIN-1, 1:1000, Sigma Life Sciences). Secondary HRP-conjugated goat anti-mouse IgG (1:20,000) or anti-rabbit IgG (1:20,000) (from Jackson ImmunoResearch, Cambridge, UK) and ECL System (Promega, Madison, WI) were used for protein detection.