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. 2020 Aug 25;39(19):e105071. doi: 10.15252/embj.2020105071

Figure EV4. Polyclonal and clonal gene targeting approaches uncover a functional CARD8 inflammasome in T cells.

Figure EV4

  • A
    MiSeq analysis of polyclonal CD4 T cells targeted either with CARD8 gRNA1, CARD8 gRNA2, or the combination of both.
  • B
    Immunoblot of VbP‐treated CD4 T cells targeted with gRNAs against the indicated genes. Two gRNAs/target gene were used. 14 days after editing and expansion, cells were treated for 18 h with VbP.
  • C
    Clonal knockout T‐cell pools of the indicated genotype were treated with VbP and morphologic changes as well as PI uptake were monitored by live‐cell imaging microscopy using a 25× objective. Fraction of PI‐pos. cells over time are shown. Images were acquired every 2 min. Background PI‐levels from the start of the experiment were subtracted. Data were normalized by a sliding window covering 30 min. Data (mean ± SEM) from three independent experiments using the same clonal knockout T‐cell pool from one donor are shown. On the right, representative images for each genotype at 8 h of VbP treatment are shown. Scale bars: 50 μm.