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. 2021 Sep 9;12:5248. doi: 10.1038/s41467-021-25523-5

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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. 2 — a–d Analysis of the (a) levels of phosphorylated (P) and/or total (T) ERK, AKT, EFR3A, and actin (as a loading control), as assessed by immunoblot, b an example of colony formation, as assessed by crystal violet staining, which is (c) plotted as the mean ± SD total colony area, and (d) the mean ± SD number of colonies growing in soft agar of human HEK-HT cells transformed by oncogenic KRAS^G12V (codon-optimized in d) engineered to stably express Cas9 and a vector encoding no sgRNA (vector) or the indicated sgRNAs. Representative of 3 biological replicates tested in 3 independent experiments. Replicate experiments and full-length gels are provided in Supplementary Fig. 11. Significance values calculated by one-way ANOVA test for (c, d): **p < 0.01. Specific p values for (c) are 0.0078 (sgEFR3A), 0.0045 (sgEFR3B), and 0.0016 (sgEFR3A/B), and (d) are 0.0059 (sgEFR3A), 0.0083 (sgEFR3B), and 0.0026 (sgEFR3A/B).