Fig. 6 | Cell Death & Differentiation

Fig. 6

From: Systematic genetic mapping of necroptosis identifies SLC39A7 as modulator of death receptor trafficking

Fig. 6

Genome-scale gene activation screens identify regulators of necroptosis. a, b Circos plots of genome-scale SAM screens in KBM7 FADD- SAM cells with necroptosis induction by TNFα (a) or SMAC mimetic birinapant (b) for 72 h. For each stimulus, screens were performed at low and high concentrations (TNFα: 10 ng/ml (green) or 100 ng/ml (purple); birinapant: 0.1 µM (blue) or 1 µM (orange)). Screen analysis was performed by identifying differentially enriched sgRNAs using DESeq2 and then aggregating sgRNAs to genes using Gene Set Enrichment Analysis. Identified hits are ranked according to the adjusted p-value of enrichment (–log10(padj). Bubble size corresponds to the average log2 fold-change (aLFC) of enrichment, color indicates the number of significantly enriched sgRNAs. Screens were performed in duplicate except the high concentration of birinapant in simplicate. c, d Cell viability in KBM7 FADD- SAM cells transduced with sgRNA targeting TNIP1, BIRC3 or Renilla luciferase (Ren). Cells were treated as indicated for 72 h and viability was assessed using a luminescence-based readout for ATP (CellTiter Glo). Data represent mean value ± s.d. of two independent experiments performed in triplicates. e KBM7 FADD- SAM cells transduced with the specified sgRNAs were lysed and subjected to immunoblotting with the indicated antibodies. Asterisk (*) indicates non-specific band

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