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. 2023 Oct 10;14:6332. doi: 10.1038/s41467-023-41828-z

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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. 3 — A H1792 and H2009 percent cell viability after Trametinib (Tram) and Midostaurin (Mido) treatment for 72 h. B Effect of Tram and Mido combination on cell viability of wild-type (wt: H1437, H2126, H1568, H1993 and H1650) and mutant (mut: H1792, H2009, A549, HCC44, H23, H358 and CP435) KRAS LUAD cells (72-h treatment). Tram: 0.5 μM; Mido: 0.625 μM (data: mean +/− SD; test: one-way ANOVA, Tukey’s adjustment). C Effect of Tram and Mido combination on cell viability of mut KRAS cells (H1792, H2009 and A549) in 3D (72-h treatment; n: 3 independent experiments). Tram: 0.01–0.05 μM; Mido: 0.05 μM (data: mean +/− SD; test: one-way ANOVA, Tukey’s adjustment). D Representative images of H1792, H2009 and A549 3D (72-h treatment). Scale bar: 200 μm. E Effect of Tram and Mido on mut KRAS patient-derived xenograft organoids (PDXOs: TP60, TP69, TP80, TP181 and TP126; 72-h drug treatment). Tram: 0.05 μM; Mido: 0.3 μM (data: mean +/− SD; test: one-way ANOVA, Tukey’s adjustment). F, G Cleaved PARP expression in mut (H1792, H2009 and A549; F) and wt KRAS (H1568, H1993 and H1437; G) cell lines after drug exposure (24-h treatment; loading control: β-TUBULIN). H Western blot of indicated proteins in H1792 and H2009 (48-h treatment; loading control: ACTIN). I Long-term assays of H1792, H2009, and A549 cells (10-day treatment; n: 3 independent experiments; data: mean +/− SD; test: one-way ANOVA, Tukey’s adjustment). Crystal violet-stained images of control, Tram- (5 nM), Mido- (100 nM) and combo-treated cells. J Percent cell viability of Tram-resistant (TR) H1792, H2009 and A549 cells (72-h treatment). K Relative depletion of shRNAs against Midostaurin targets in Trametinib-treated versus doxycycline-treated H23 cells (n: 6 shRNAs/gene). Blue: hit selected by Manchado et al.⁷. Red: hits sensitizing to Trametinib with >30% relative depletion. Orange: hits sensitizing between 20% and 30%. Brown: hits sensitizing between 10% and 20%. L Effects of Tram (10–50 nM) and Barasertib (Bara; 25–1000 nM) combination on cell viability of mut KRAS (H1792, H2009, A549, HCC44, H23, H358) cells (72-h treatment; data: mean +/− SD; test: Kruskal–Wallis, Dunn’s adjustment).