Extended Data Fig. 1: Validation of high-throughput tethering approach.
From: Deciphering disordered regions controlling mRNA decay in high-throughput
a,b, Gating schemes used for flow cytometry analysis. These criteria were applied across all subsequent analyses. c, Representative raw YFP and RFP distributions for tethering controls. d, Mean iRFP/RFP ratios from flow cytometry for tethering fragments. Individual values from each replicate and standard deviation are displayed (n = 3). e, Normalized read distribution of two fragments across bins. Colour shades show two independent biological replicates. f, Gating scheme for FACS on the iRFP/RFP distribution. Bins divide the cells into four roughly equal-sized populations. g,h, Correlation between replicates for g, activity scores and h, stability scores, with Pearson correlation coefficient displayed in figure panels. i, Distribution of RFP signal from mScarlet normalizer for each replicate measured in Fig. 1f. j, Plasmid shuffling assay of Nab3 variants in media lacking uracil (Left) or containing 5-FOA (right).
