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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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1D analysis of Hsp90. (A) Movement of Hsp90 along the single degree of freedom (CMA). The rotation of chain A relative to a fixed chain B. (B) Examples of the synthetic images with varying SNR between [0.001, 0.1]. (C) Free-energy profiles along the path for the entire set of images recovered from cryo-BIFE. The ground truth free-energy profile is shown in black. The expected free energy profile using cryo-BIFE is shown for BioEM orientation rounds 1 and 2 in orange and blue, respectively. The R-hat test for the MCMC stationarity yielded 1.000 and 1.001 for BioEM round 1 and 2, respectively. The bars show the credible interval at 5% and 95% of the empirical quantile at each node. A cubic spline is used to fit the expected free-energy profile, providing a smooth profile.