Fig. 2: DNA damage induces interactions between shieldin and CST.
a, Schematic depicting shieldin purification and proteomic elucidation strategy. Briefly, Shld1/3-knockout CH12-F3 cell lines were lentivirally transduced with TwinStrep–HA–Shld1/3 espression transgenes. First, 300-ml suspension cultures were either mock-treated or irradiated (10 Gy) and lysates were prepared following a 1-h recovery. Shieldin complexes captured from lysates on MagStrep-XT resin were eluted in biotin, captured and tryptic-digested on S-Trap columns, with resulting peptides analyzed by LC–MS/MS. The schematic was generated using BioRender.com. b, B cell SHLD3 versus control (beads only) interactomes as defined by LC–MS/MS and LFQ. The scatter plot depicts the log2 fold enrichment of indicated MagStrep-XT purified complexes across two independent experiments. c, Comparison of mock and irradiated B cell SHLD3 interactomes as defined by LC–MS/MS and LFQ. The scatter plot depicts the log2 fold enrichment of indicated MagStrep-XT purified complexes across two independent experiments. d, Immunoblot analysis of CH12-F3 cell extracts from SHLD3 pulldown experiments. Representative of n = 2 independent experiments.
