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. 2023 Dec 22;120(52):e2310542120. doi: 10.1073/pnas.2310542120

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Copyright © 2023 the Author(s). Published by PNAS.

This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 2. — Mutation of S70F affects the phase separation of HEI10. (A) Immunostaining of HEI10 at pachytene, diplotene, and diakinesis in Col-0, HEI10-Flag, and HEI10^S70F-Flag transgenic plants. (Scale bar, 5 μm.) (B) Immunostaining of HEI10-Flag and HEI10^S70F/A/D-Flag in tobacco nuclei with or without MMS treatment. The leaves are treated with MMS for 1.5 h. HEI10^S70A and HEI10^S70D simulate the phospho-dead and phospho-mimetic proteins. (Scale bar, 5 μm.) (C) In vitro phase separation assay of mRFP, mRFP-HEI10, and mRFP-HEI10^S70F recombinant proteins, with protein concentrations of 15 μM and 10% PEG8000. (Scale bar, 20 μm.) (D) The 3D structures of HEI10 (green) and HEI10^S70F (blue), predicted by ESMFold. The Ser70 (yellow) and the Phe70 (purple) residues are shown at the junction between Ring domain and α-helix by stick representation.