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. 2020 Jan 20;10:635. doi: 10.1038/s41598-020-57500-1

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© The Author(s) 2020

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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Gene repression and activation at the transcript and protein levels. (a) RT-qPCR analysis of TCF4 3b transcript levels at indicated time points in dCas9-KRAB-PB clones K1 and K2 separately and averaged. Data is shown as mean +/− s.e.m. *p = 0.0074, **p = 0.0017. (b) Western blot analysis of TCF4 protein in dCas9-KRAB-PB clones K1 and K2 in biological triplicate at indicated time-points with quantification shown on the right. *p = 0.0003, **p = 0.0013 and ***p = 0.0324. (c) RT-qPCR analysis of TCF4 3b transcript levels at indicated time points in dCas9-VPR-PB clones V1 and V2 separately and averaged. Data is shown as mean +/− s.e.m. *p = 0.0210, **p = 0.0086. (d) Western blot analysis of TCF4 protein in dCas9-VPR-PB clones V1 and V2 in biological triplicate at indicated time-points with quantification shown on the right. *p = 0.0037, **p = 0.02901, ***p = 0.0082. (e) Overview of single-gRNA PB vector cloning, delivery, and selection. (f) Western blot analysis of TCF4 protein with single gRNAs in dCas9-KRAB and dCas9-VPR cells in biological triplicate with quantification shown on the right. *p = 0.0265. (g) Western blot analysis of POGZ protein with single gRNAs in dCas9-KRAB and dCas9-VPR cells in biological triplicate with quantification shown on the right. *p = 0.0097. In all Western blot panels, expression levels are normalized to GAPDH.