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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. 7 — a–d Immunoblot analysis of (a) membrane fraction for myc-KRAS^G12V, myr-myc-KRAS^G12V, C-RAF, Na+/K+ ATPase, and GAPDH, and (b) phosphorylated (P) and total (T) ERK and AKT, and (c) an example of colony formation, as assessed by crystal violet staining, which is (d) plotted as mean ± SD of colony area, for human HEK-HT cells stably infected with a lentivirus encoding Cas9 encoding no sgRNA (vector) or sgEFR3A/B in the absence and presence of ectopic myc-KRAS^G12V or when targeted to the PM by an N-terminal myristoylation sequence (myr-myc-KRAS^G12V). INPUT levels of indicated proteins serve as loading controls. a, b Representative of 3 biological replicates. c, d Representative of 3 biological replicates tested in triplicate. Replicate experiments and full-length gels are provided in Supplementary Fig. 13. Significance values were calculated by one-sided student’s t test for (d): ***p < 0.001. Specific p values for (d) are 0.00027 (KRAS^G12VsgEFR3A/B) and 0.00022 (myr-KRAS^G12VsgEFR3A/B).