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. 2020 Feb 12;9(1):1724049. doi: 10.1080/2162402X.2020.1724049

Figure 4.

Figure 4.

Characterization of AZD8055-sensitive and -resistant mILCs by immunohistochemistry and RPPA analysis. (a) Schematic overview of experimental setup to generate the different tumor groups. Mice were transplanted orthotopically with a 1 mm3 fragment of a mILC from a KEP donor mouse. When tumors reached a diameter of 5 mm, daily treatment with AZD8055 (green arrows) or vehicle control solution (black arrows) was started. AZD8055-sensitive tumors (n = 10) were harvested after 5 days of AZD8055 treatment. AZD8055-resistant tumors (n = 20) were harvested when they progressed on AZD8055 treatment to a diameter of 15 mm. Vehicle-treated control tumors were harvested after 5 days (n = 4) or when they reached a diameter of 15 mm (n = 6). (b) Immunohistochemical quantification of percentages of Ki-67 positive tumor cells in peripheral tumor parts. * p < .05; *** p < .001. (c) Immunohistochemical quantification of number of cleaved caspase 3 (CC3) positive tumor cells per 10 high magnification fields of view. (d) Unsupervised hierarchical clustering analysis of Reverse Phase Protein Array (RPPA) data from 29 KEP tumors (9 control tumors, 5 AZD8055-sensitive tumors, and 15 AZD8055-resistant tumors). The heatmap shows expression levels of selected epitopes representing known PI3K signaling markers. The complete heatmap is shown in Supplementary Fig. S4.