Extended Data Fig. 5: Analysis of E3 activity in proximity-dependent degradation.
From: Proteome-scale discovery of protein degradation and stabilization effectors
(a) Comparison of two independent replicates for the degradation assay with 290 individual vhhGFP-tagged E3s transfected into the EGFP-ABI1 293T reporter cell line. (b) Relative mRNA expression of EGFP in 293T EGFP-ABI1 reporter cell line transfected with indicated effectors fused to vhhGFP. GAPDH mRNA levels were used to normalize the EGFP mRNA values. (c) Comparison of degrader hits recovered from at least one of the pooled ORFeome screens (vhhGFP or PYL1) and non-hits in arrayed degradation assay. Statistical significance was calculated with unpaired two-tailed Mann Whitney test (**, p < 0.0036). (d) Effect of different E3 families on EGFP-ABI1 reporter stability in 293T cells. RING, Really Interesting New Gene; BTB (also known as POZ), BR-C, Ttk and Bab; CRL, Cullin E3 RING ubiquitin ligase; DCAF, DDB1 and Cul4 associated factor. (e) Phylogenetic clustering of FBXL family E3s and their activity in the degradation assay in the EGFP-ABI1 293T reporter cell line. (f) As in D, but for BTB-BACK-Kelch family E3s. (g) Comparison of the activity of full-length E3 constructs (FL) to their splice variants without the RING domain in the EGFP-ABI1 293T reporter cell line.
