Extended Data Fig. 1: Establishing the screening system.
(a) Schematic of the GAL4-based transactivation screening platform. ADs of interest are cloned downstream of the GAL4 DNA binding domain and recruit their respective cofactors to activate a reporter containing a fast-degrading GFP. Both constructs are lentivirally integrated. (b) Alpha-fold predictions of the structure of the 9 transcription factors used in the CRISPR screens. The specific AD region used is highlighted in green. (c) Flow cytometry analysis of demonstrating loss of GFP prior to cell death based on FSC-A and SSC-A and GFP signal at different timepoints after addition of triptolide (10uM). (d) Quantification of the reduction in GFP signal (average fluorescent signal, M.F.I) at different timepoints after the addition of triptolide (10uM). (e) Flow cytometry analysis of GFP signal at day 5 after infection with indicated sgRNAs in GAL4-VP64 and GAL4-MYB AD lines. FSC-A and SSC-A plots shown below to demonstrate predominantly live cells at the timepoint analysed. All flow analysis is performed on at least 10000 cells.
