Extended Data Fig. 2: Characterization of DMS Libraries and AT7.
From: Functional E3 ligase hotspots and resistance mechanisms to small-molecule degraders
(a) Pie charts depicting the distribution of different alterations identified by sequencing the mutational scanning libraries for CRBN (top) and VHL (bottom). (b) Stacked bar graphs and density distributions of residue wise normalized abundance of mutants identified in the DMS libraries for VHL (top) and CRBN (bottom). (c) Chemical structure comparison of the degraders AT1 and AT7. (d) Dose-resolved, normalized viability after 3 d treatment with MZ-1, macroPROTAC-1, cis MZ-1 (a non VHL binding control of MZ-1 or AT7 in MV4;11 cells. Mean ± s.e.m.; n = 3 independent treatments. (e) Dose-resolved, normalized viability after 3 d treatment (AT7) in RKO VHL−/− cells with over-expression of VHLWT. Mean ± s.e.m.; n = 3 independent treatments. (f) Protein levels in HeLa cells treated with MZ-1 or AT7 (18 h, indicated concentration). Representative images of n = 2 independent measurements. (g) Protein levels in RKO VHL−/− cells with over-expression of VHLWT treated with DMSO or AT7 (60 nM, 2 h). Representative images of n = 2 independent measurements. (h) Dose-resolved, normalized viability after 4 d treatment (ACBI1) and 3 d treatment (ARV-771, MZ-1, macroPROTAC-1) in RKO VHL−/− cells with over-expression of VHLWT. Mean ± s.e.m.; n = 3 independent treatments.
