Fig. 1

CRISPR screen uncovers an essential role for ERK2 in IFNγ-mediated growth inhibition. (A) Melanoma lines were treated with or without IFNγ in triplicate, and cell growth was monitored for 7 days in an IncuCyte live imaging experiment. Line names are color-coded to indicate melanoma subtypes: Maroon, BRAF mutant; Green, NRAS mutant; Cyan, NF1 mutant; Orange, triple wild type; Red, BRAF and NRAS double mutant; Dark blue, BRAF and NF1 double mutant; Black, not determined. Refer to Supplementary Table 2 for cell line details. Growth curves for selected lines are presented in Fig. S1. (B) Schematic depicting the experimental setup for all three CRISPR screens. (C) Representative volcano plot with results from the GeCKO library. Fold enrichment or depletion in the IFNγ-treated sample calculated over all sgRNAs targeting each gene is shown on the x-axis, while the y-axis shows the statistical significance for each gene. Top 5 enriched genes are labeled on the plot. (D) Lists of the top 30 genes enriched in the IFNγ-treated sample for all three screens were compared to identify top hits common to all three screens. (E) Plot showing fold enrichment values for all the individual sgRNAs targeting ERK2 (MAPK1) and genes involved in the core IFNγ signaling pathway from each screen. ERK2 is enriched to a similar level as the IFNγ signaling genes across all three screens. See also Fig. S2