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. 2018 Mar 19;9:1138. doi: 10.1038/s41467-018-03503-6

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© The Author(s) 2018

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. 3 — The lower helical boundary of CRY’s secondary pocket gates interaction with CLOCK. a A surface view of the structure of CRY1 (PDB: 5T5X) with the lower helix of the secondary pocket shown as a ribbon diagram with side chains shown as sticks and relevant residues color-coded for the rest of the figure. Inset is a magnified view of this helix with residues labeled. b Schematic of Cry1 rescue assay. RRE is an ROR response element necessary for delayed expression of CRY. c Rescue assays with CRY1 mutants identified in the SCA (n = 3 per condition, reflective of 6 plates from two independent experiments) shown as means ± SEM. d Co-IP assay with PER2, CLOCK, BMAL1, and various CRY1 mutants. Multiple bands in PER2, CLOCK, and BMAL1 lanes are indicative of post-translational modifications on these proteins. Blot is representative of at least three independent experiments. e Rescue assays for two CRY1 residues not identified in the SCA (n = 3 per condition, reflective of 6 plates from two independent experiments) shown as means ± SEM. f Period plot for the data shown in e. Asterisks show significance by unpaired t test with Welch’s correction (**p = 0.0018, ****p < 0.0001)