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. 2023 Oct 30;14:6929. doi: 10.1038/s41467-023-42643-2

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© The Author(s) 2023

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 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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Fig. 4 — a MS2 system for visualization of transcription in single living cells. Transcription of the 24xMS2 DNA array generates RNA stem loops that are detected through their recruitment of Halo-tagged coat proteins and visible as fluorescent spots at the transcription site (b). CDS: Coding sequence. b Oct4-MS2 live-imaging reporter in WT mESCs shows transcriptional bursting. Shown are example microscopy data for indicated time points (dashed line) of the time course. Scale bar: 2.5 µm. c Oct4-MS2 signal as a function of steady-state nuclear LEXY-YAP levels reveals that YAP acts as a repressor of Oct4 transcription. Min-max range of endogenous YAP levels measured in WT cells is indicated by gray shading. Shown are mean ± SEM, N = 15 independent experiments. d Top: Oct4-MS2 transcriptional activity upon light-gated nuclear YAP export in LEXY-YAP mESCs (red curve) vs. non-light responsive YAP control cells (gray curve). Difference between mean of LEXY-YAP and YAP curve is shown as inset on the top right, indicative of transient adaptive YAP target activation in response to sustained export of YAP from the nucleus. Bottom: Nuclear YAP levels in LEXY-YAP (red) and YAP (gray) mESCs simultaneously imaged for Oct4-MS2 shown in the top panel. Illumination phase is indicated by blue shading. Shown are mean ± SEM, N = 12 independent experiments.