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. 2019 Mar 4;26(11):2400–2415. doi: 10.1038/s41418-019-0309-6

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© The Author(s) 2019

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 — Targeting of KRAS synergizes with trametinib in suppressing the viability of KRAS mutant colon cancer cells in vitro and in vivo. a Cell proliferation assays using the Cell Counting Kit-8 in SW1116 and SW480 cells transfected with ΔN-KA-EE-MEK or control vector, in the presence or absence of YAP silencing at day 3. b Cell proliferation assays at day 3 of SW1116 and SW480 cells cultured with deltarasin (1M) or DMSO in the presence or absence of 100 nM trametinib. The data presented as the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test (two-tailed). c Clonogenic assays of SW480 and SW1116 cells cultured with DMSO or 1 µM deltarasin (or/and 100 nM trametinib) at day 7. d Cell proliferation assays in SW1116 and SW480 cells transfected with siKRAS or negative control, and then cultured with 100 nM trametinib or DMSO for 3 days. The data presented as the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test (two-tailed). e Tumour formation assays in the nude mice subcutaneously injected with SW480^Teto-shKRAS (group shKRAS and shKRAS + tram) and SW480^{Teto-shControl}. The mice were treated with DMSO (group Control and shKRAS), deltarasin (15 mg/kg), trametinib (3 mg/kg) in the vehicle (20% PEG300, 5% Tween 80 and normal saline) according to groups via intraperitoneal injection daily. All groups were fed with doxycycline water (2 g/L). The bar graph indicates the tumour weight of each group (n = 5). The tumour sizes were measured every 5 days. The line charts show the tumour volume of each group (n = 5). The data presented as the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test (two-tailed). f Representative pictures of MEK staining in xenograft tumour tissues. The graph indicates the IHC scores of nuclear MEK expression, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test (two-tailed). g Representative pictures of YAP staining in xenograft tumour tissues. The graph indicates the IHC scores of nuclear YAP expression, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test (two-tailed). h Quantitative real-time RT–PCR to measure YAP mRNA level in xenograft tumour tissues. GAPDH was used as a control, ***p < 0.001 using Student’s t test (two-tailed)