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. 2021 Jul 1;11:13657. doi: 10.1038/s41598-021-92621-1

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© The Author(s) 2021

Open AccessThis 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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2D analysis of Hsp90. (A) Two degrees of freedom of Hsp90 along the CMA and CMB rotation directions (see the Methods). (B) Ground truth free-energy surface along CMA and CMB directions. Black (CV1), orange (CV2) and green (CV3) dashed lines show three paths used for the cryo-BIFE analysis. (C) The free-energy profiles along these three path CVs, extracted with cryo-BIFE using synthetic particle images (dashed lines), are compared to the ground truth projected profiles (solid lines). The R-hat test for the MCMC yielded values $< 1.003$ for all cases. The bars show the credible interval at 5% and 95% of the empirical quantile at each node. The results are for the second BioEM round of orientation estimate.