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. 2021 Mar 2;40(12):2309–2322. doi: 10.1038/s41388-021-01708-6

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© The Author(s) 2021

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. 4 — A Scheme showing the CRISPR-Cas9 screening strategy to identify miR-200b/c regulators using the miR-200b/c sensor. B Graphs showing read counts for the indicated genes’ gRNAs between sorted miR-200b/c high and control cells. Validation of the top hits from the CRISPR/Cas9 screen by qPCR. miR-200b (C) and miR-200c (D) levels were quantified in cells stably transfected with the cDNA clones overexpressing the hits genes, compared to the pCDNA3.1 vector control. E Western blot quantification of EMT markers ZEB1 and E-Cadherin in cells as in (C, D). ZEB1 and E-Cadherin band intensities were calculated by densitometry. β-Actin was used as a loading control. p values are from Student’s t-test. Points are average ± SD. *<0.05, **<0.01, ***<0.001, ****<0.0001.