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. 2022 Jul 21;54(8):1238–1247. doi: 10.1038/s41588-022-01132-w

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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 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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Extended Data Fig. 7 — a. IAP RNA facilitates RNAPII CTD droplet formation in vitro. Displayed are representative images of droplet formation by purified RNAPII CTD-mCherry fusion proteins in the presence of in vitro transcribed IAP RNA fragments. Scale bar: 10 μm. b. IAP RNA facilitates MED1 IDR droplet formation in vitro. Displayed are representative images of droplet formation by purified MED1 IDR-mCherry fusion proteins in the presence of in vitro-transcribed IAP RNA. c. Partitioning ratio of MED1 IDR-mCherry into droplets at the indicated IAP RNA concentrations. Every dot represents a detected droplet. Data for the quantification was acquired from at least five images of two independent image series per condition. P value is a from a two-tailed t test. d. IAP RNA is enriched within MED1 IDR droplets. Displayed are representative images of the enrichment of fluorescein-labeled IAP RNA in MED1 IDR-mCherry droplets. e. Quantification of the enrichment of fluorescein-labeled IAP RNA in MED1 IDR-mCherry droplets. Data for the quantification was acquired from at least five images of two independent image series per condition. P value is from a two-tailed t test. f. (left) Schematic model of the heterotrimeric NFY transcription factor. (right) Graphs plotting intrinsic disorder in the NFYA, NFYB and NFYC proteins. The NFYC-IDR cloned for subsequent experiments is highlighted with a blue bar. g. Concentration-dependent droplet formation by purified recombinant NFYC IDR-mEGFP. Scale bar: 10 μm. h. Representative images of droplet formation by purified NFYC IDR-mEGFP and MED1 IDR-mCherry fusion proteins. MED1 IDR-mCherry mixed with purified mGFP is included as a control. Scale bar: 10 μm. i. Partitioning ratio of NFYC IDR-mEGFP or mEGFP in MED1 IDR-mCherry droplets. Every dot represents a detected droplet. Data for the quantification was acquired from at least five images of two independent image series per condition. P value is a from a two-tailed t test.