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. 2020 Jun 19;39(31):5373–5389. doi: 10.1038/s41388-020-1364-7

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© The Author(s) 2020

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. 6 — a Sphere formation assay on MiaPaCa2 cells expressing either control shRNA or ICMT-targeting shRNA, with concurrent expression of CA-KRAS, the PI3K catalytic subunit p110, the kinase active RAF-22W, or the empty vector control as indicated. The images of the spheres formed are shown on the left side; the quantifications of the spheres, using OpenCFU and Prism5 software on three technical repeats, are plotted on the right side. b The same experiment was performed as shown in a but with MDA-MB231 cells. c, d Immunoblot analysis of the same cells used for a and b to show the TAZ levels and the engagement of pMEK downstream of RAF and pAKT downstream of p110, as the result of the expression of CA-KRAS, RAF-22W, or p110. e Sphere formation assays on MiaPaCa2 and MDA-MB231 cells treated with increasing concentrations of MEK inhibitor PD184352. The images of the spheres formed are shown on the left side; the quantification to calculate the means and standard deviations, using OpenCFU and Prism5 software, on three technical repeats, are plotted on the right side. f Immunoblot analysis for TAZ and pERK levels in MiaPaCa2 and MDA-MB231 cells treated with the indicated concentrations of PD184352. All studies have been repeated three times with similar results.