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. 2022 Oct 27;12:18091. doi: 10.1038/s41598-022-22788-8

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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T cell activation induces extracellular Hsp90 (mHsp90) expression. (a) Chemical structure of HS-131, and HS-132 active molecule, the inactive analogue HS-198 (negative control). (b) Naïve CD3+ T cells isolated from healthy human peripheral blood mononuclear cells (PBMC’s) were activated with anti-CD28 and anti-CD3 antibodies, or resting (unstimulated) for 96 h and treated with HS-132 at the indicated concentrations. Data represented as mean ± SD, 6 biological replicates. (c) Internalization of probe by CD3+ T cells at physiological (37 °C) and non-physiological (4 °C) temperatures at varying concentrations of HS-132. Data represents mean ± SD, 4 biological replicates. (d) CD3+ T cells isolated from stimulated human PBMC’s were treated with 10 µM HS-131 (active molecule) and HS-198 (inactive control) ± a tenfold excess of ganetespib. Data represents mean ± SD, 3 biological replicates. (e) Confocal analysis of HS-132 uptake in stimulated CD3+ T cells, which is blocked with treatment of the non-fluorochrome tethered Hsp90 inhibitor HS-10.