Fig. 4 | Cell Death & Differentiation

Fig. 4

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

Fig. 4

Orthogonal genetic screens and surface marker analysis specify SLC39A7 loss-of-function phenotype on receptor trafficking. a Spider plot summarizing the results of 8 independent CRISPR/Cas9 screens in KBM7 wildtype, KBM7 FADD-, or HAP1 cells using an SLC-specific gRNA library. Each plot section represents one screen with the indicated stimuli. 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 indicates the number of significantly enriched sgRNAs and color the average log2 fold-change (aLFC) of the enriched sgRNAs. Screens were performed in duplicate. SLC39A7 is highlighted in orange. No gene was identified in KBM7 wildtype cells upon TRAIL treatment. b MCA of Jurkat E6.1 SpCas9 cells transduced with a GFP marker (GFP+) and sgRNAs targeting either SLC39A7 or Renilla luciferase (sgRen) as control, against cells transduced with sgRen and an mCherry marker (mCherry+). The cell populations were mixed at 1:1 ratio, treated with TRAIL (20 ng/ml) or FASL (1 ng/ml), and analyzed after 14 days by flow cytometry. Data represent mean value ± s.d. of two independent experiments performed in duplicates, n.d. (not determined) indicates wells with no outgrowth. c Flow cytometry analysis for TRAILR1 (left) and TRAILR2 (right) surface expression in KBM7 FADD-, KBM7 FADD- sgRen or KBM7 FADD- SLC39A7- cells. Data shown are representative of two independent experiments

Back to article page