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. 2023 Sep 20;14:5856. doi: 10.1038/s41467-023-41583-1

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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. 7 — a Representative WB images of lysates from KPB6 control cells expressing Cas9 (CRISPR^Cas9) and from derived, SOS1-silenced (CRISPR^SOS1) and SOS2-silenced (CRISPR^SOS2) cells, immunolabelled for SOS1, SOS2 or tubulin. The bar graph quantitates the relative expression level of SOS1 or SOS2 normalized to that of tubulin in each case. n = 3 independent samples per group. Data shown as mean ± SD. ****P < 0.0001 vs CRISPR^SOS1/2-depleted KPB6 cells. One-way ANOVA and Tukey’s test. b Graph displaying growth curve of KPB6 cell cultures at 24, 48, and 72 h upon CRISPR-induced SOS1 or SOS2 ablation. n = 12 independent samples for CRISPR^Cas9 and n = 8 independent samples for CRISPR^SOS1/2 groups. Data shown as mean ± SD. One-way ANOVA and Tukey’s test. At 24 h (*P = 0.00379 vs CRISPR^SOS1 and ^#P = 0.00241 vs CRISPR^SOS2); at 48 h (**P = 0.0020 vs CRISPR^SOS1 and ^##P = 0.0013 vs CRISPR^SOS2) and at 72 h (**P = 0.0022 vs CRISPR^SOS1 and ^##P = 0.0078 vs CRISPR^SOS2). c Relative levels of RAS-GTP in cellular extracts from steady-state cultures of KPB6 cells upon CRISPR-mediated SOS1 or SOS2 depletion. n = 6 independent samples per group. Data shown as mean ± SD. **P = 0.0038 vs CRISPR^SOS1. One-way ANOVA and Tukey’s test. d Proliferative rates of cultures of the indicated KRAS^G12D-mutated cell lines upon treatment with DMSO (vehicle), BAY-293, MRTX1133 or combo (BAY-293 + MRTX1133) for the times indicated. n = 5 independent samples per group. Data shown as mean ± SD. ****P < 0.0001 vs drug-treated cells. One-way ANOVA and Tukey’s test. e Relative levels of RAS-GTP in extracts from serum-starved cultures of the indicated cell lines (white boxes) as well as EGF-stimulated cultures of the same cell lines after undergoing pretreatment with solvent vehicle (blue) or the indicated drugs, singly or in combination. n = 3 independent samples per group. Data shown as mean ± SD. *P < 0.05 (P = 0.0498 in LKR13 cells) or ***P < 0.001 (in KPB6 cells, P = 0.0009 and P = 0.0008, for MRTX + EGF or combo-treated groups, respectively) vs vehicle-treated, EGF-stimulated group, and ^#P < 0.05 (P = 0.0354 and P = 0.0316, for MRTX + EGF or combo-treated groups, respectively) vs BAY293 + EGF group. One-way ANOVA and Tukey’s test. Source data are provided as a Source data file.