Extended Data Fig. 2: Reproducibility and validity of AD screening system.
(a) Quantification of change in GFP at day 5 after infection with sgRNAs targeting indicated cofactors in two independent AAVS reporter knock-in clones activated by different ADs. CDK8i was applied at a dose of 10uM for 24 hrs. Change in GFP in polyclonal reporter lines provided for direct comparison. Analysis of polyclonal lines was also performed 5 days after infection with sgRNAs targeting indicated cofactors. Error bars represent S.E.M of two independent sgRNAs or drug treatments. (b) Correlation plot of the fold enrichment between two independent biological replicates of the GAL4-NF-kB mCMV screens. Each replicate was performed using an independently derived reporter line likely to contain different lentiviral integration sites. r = Pearson correlation. Error bands reflect 95% confidence interval of Pearson correlation. (c) Correlation plot of the average fold enrichment scores between two independent biological replicates of VP64-AD screens. One replicate was performed using the GAL4-VP64 and a lentiviral reporter. The second replicate was performed using ZFP-VP64 and an insulated piggyBac reporter. r = Pearson correlation. Error bands reflect 95% confidence interval of Pearson correlation. (d) Flow cytometry analysis of GFP signal in each of the GAL4-AD cell lines at day 5 after infection with control sgRNAs and two independent sgRNAs targeting GAL4.
