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. 2021 Dec 13;4:1391. doi: 10.1038/s42003-021-02906-4

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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. 5 — a Coomassie blue staining (CBS) showing proteins trapped by biotin-labeled D6 (D6-4) from NCI-H1975 cell lysates. 10 × D6: using 10-fold D6 to saturate the target proteins and thus excluding non-specific binding. The protein gel indicated by dashed line was further analyzed by protein mass spectrometry and identified proteins are listed in the right panel. b, c Immunoblotting analysis of the protein elution based on biotin–avidin pulldown assays from NCI-H1975 cell lysates (b) or purified recombinant GST-HSP90 protein (c); PDS: pulldown solution. d Immunoblotting analysis showing the domains where D6 binds to HSP90. e Immunoblotting analysis of the cell lysates from A549, PC9, and NCI-H1975 cells treated with D6 (2.5 μM) or 17-AAG (2.5 μM), respectively. f Immunoblotting analysis of the anti-HSP90 immunoprecipitates prepared from cell lysates of NCI-H1975 cells with indicated concentrations of D6 treatment. g Immunoblotting analysis of anti-HSP90 immunoprecipitates derived from cell lysates of HEK293 cells transfected with HSP90 and indicated EGFR mutants with or without D6 treatment.