Fig. 3: Application of RADAR to pooled CRISPR screening.

A Schematic of how the pooled CRISPR screen was performed. B GFP levels for cells transduced with the CP1 gRNA library and treated with DMSO or ABA. C Comparison of RADAR-expressing cell activity (as measured in GFP levels) when treated with or without CP1 gRNA library. D RSA-down and RSA-up values for genes in CP1 and CP3 gRNA libraries, plotted against maximum log₂(fold change). Positive regulator gene hits scoring −4 and below are highlighted in red, with example names displayed for genes associated with the PKC-induced AP-1 pathway. Negative regulator gene hits scoring −4 and below are displayed separately for ABA (orange) and ABA+PMA (green) conditions. E Robust z-scores of reporter activity, for genes included in the reconfirmation screen. Activity for RADAR was measured in terms of %GFP− cells for positive regulator screen, and %GFP+ cells for negative regulator screen. Activity for classical reporter was measured by GFP mean values for both positive and negative regulator screens. Dotted line indicates 3 standard deviations away from the median negative control activity (n = 3, mean ± s.d.). F Flow cytometry data for cells expressing either reporter, and with or without gRNA targeting SATB2. RADAR and classical reporter-expressing cells were treated with ABA/PMA or PMA, respectively, to activate the AP-1 pathway, then GFP levels measured.