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. 2021 Nov 25;12:6896. doi: 10.1038/s41467-021-27210-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a The structure of the BCL-xL and/or BCL-2 degraders (racemic mixture). b The BCL-xL and BCL-2 degradation profiling of the PROTACs evaluated in 293T cells after the cells were treated with four different concentrations of indicated PROTACs for 16 h. The DC₅₀ and D_max were calculated using Prism based on the ImageJ-quantified western blots in Supplementary Fig. 4. c, d AlphaLISA assay to determine the formation of BCL-xL/PROTAC/VHL-ELOC-ELOB (VCB) (c) or BCL-2/PROTAC/VCB (d) ternary complex in a cell-free condition. Data represent the mean of a single experiment with 2 technical replicates. Similar results were obtained in one more independent experiment. e, f NanoBRET assays evaluating intracellular BCL-xL/PROTAC/VCB (e) or BCL-2/PROTAC/VCB (f) ternary complex formation. 293T cells were transiently transfected with HiBit-BCL-xL, LgBit, and HaloTag-VHL or HiBit-BCL-2, LgBit, and HaloTag-VHL and then treated with a serial dilution of the indicated compounds for 6 h. Data are expressed as mean ± s.e.m. of three biological replicates. Source data are provided as a Source data file.