Extended Data Fig. 7: Mass spectrometry-based analysis of SIGLEC12 phosphorylation during necroptosis and its effect of SIGLEC12-mediated PMR.
From: SIGLEC12 mediates plasma membrane rupture during necroptotic cell death
a, A large-scale IP-FLAG of SIGLEC12 from HT-29-SIGLEC12-KO-SIGLEC12-FLAG stable cell line following indicated treatments was resolved on SDS-PAGE and stained with Coomassie brilliant blue. Boxes indicate the bands that were analyzed by mass spectrometry to perform phosphomapping. LC-MS/MS ion fragmentation graphs of peptides containing phospho-T519, phospho-S532/S539, and phospho-T555, respectively. pT and pS indicate the phosphorylated residues. b, 293FT-SIGLEC12-FL-WT, 293FT-SIGLEC12-FL-4TS/A, and 293FT-MLKLQ356A cells were treated with 0.1 μg/ml doxycycline, and cell death was quantified using Incucyte S3. Data were plotted as the mean ± s.d., representative of n = 4 (b); three independent experiments. Data are representative of three independent experiments (a, b).
