Anti fas c 20

TNF-α and z-VAD-fmk were purchased from R&D. Cyclohexamide (CHX) from Sigma. Smac mimetic was a gift from S. Wang (University of Michigan, Ann Arbor, Michigan, USA). TRAIL from Novoprotein. FasL from Upstate Biotech. CH11 from MBL Medical and Biol Lab.
All antibodies are at concentration of 1 μg ml⁻¹ and used at 1:1,000 dilution unless otherwise stated. Anti-RARγ (C-15) (sc-550) for human, anti-RARα (C-20) (sc-551), anti-caspase-8 (C-20) (sc-6136), anti-cIAP2 (sc7944) and anti-Fas (C-20) (sc-715) from Santa Cruz; anti-RIP1 (610459) and anti-FADD (610400) from BD Biosciences; anti-RARγ1 (ab5905) for mouse, anti-RIP3 (ab72106), anti-MLKL (ab184718) for human, anti-MLKL (ab172868) for mouse and anti-cIAP1 (ab2399) from Abcam; anti-RIP3 (2283) for mouse from ProSci, anti-TRADD (05-473) from Upstate; anti-TRAF2 (MAB3277) and anti-TNFR1 (AF-425-PB) from R&D; anti-Actin (A3853) (dilution 1:10,000), anti-FLAG (F9291) (dilution 1:5,000) and anti-GFP (G6539) (dilution 1:5 000) from Sigma; anti-V5 (R960-25) (dilution 1:5,000) from Invitrogen; anti-PARP1 (BML-SA250-0050) from Enzo Life Science; anti-GAPDH (NB300-22) (dilution 1:5,000) from Novus Biologicals; anti-cleaved caspase-8 (9496), anti-CYLD (4495), anti-RIP1 (137451) and anti-p-RIP1(65746) from Cell Signaling Technology; anti-DsRed (632392) (dilution 1:5,000) from Clontech.

Western blot: anti-CD40 S-17 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-Fas (ZB4) (Abcam, Cambridge, UK), anti-DR5 (Millipore, Billerica, MA, USA). Immunoprecipitations: anti-CD40 C20 agarose conjugate (Santa Cruz Biotechnology), anti-Fas (C20) (Santa Cruz Biotechnology), anti-DR5 D4E9 (Cell Signalling, Danvers, MA, USA). Flow cytometry: anti-CD40-FITC and –PE-Cy5 (5C3), anti-Fas-PE (DX2), anti-MHC-II-FITC (Tü 39), anti-CD86-PE (2331) and anti-CD19-V500 (HIB19) (BD Pharmingen, San Diego, CA, USA); anti-CD27-BV421, anti-CD95-AF647 and anti-IgD-PE-Cy7 (Biolegend, San Diego, CA, USA); anti-DR5-PE (B-K29) (Diaclone Research, San Diego, CA, USA) for cell lines; anti-TRAILR2 PE (eBioscience, San Diego, CA, USA) for primary cells; anti-TRAILR2 (TR2.21; to screen TRAILR2 KO BJAB clones) (Adipogen, Lausen, Switzerland); anti-phospho-NF-κB p65 (Ser536, clone 93H1)-AF647 (Cell Signalling). Flag-ACRP-hBAFF, Flag-ACRP-hCD40L and Flag-ACRP-FasL were produced in house and staining were performed as described in Bossen et al.^{27 (link)} Flag-ACRP-hCD40L (mega-CD40L) was obtained from Adipogen. Anti-TRAILR3 (LEIA) was used for GPI recognition (own production).^{27 (link)} Microscopy and PLA: monoclonal rabbit anti-CD40 (EBI-19-21) (Abcam), mouse anti-DR5 (DJR2-4) (ABD Serotec, Raleigh, NC, USA) and mouse anti-CD95 (DX2) (BD Biosciences, San Diego, CA, USA).