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. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Cancer Lett. 2018 Jun 20;433:1–9. doi: 10.1016/j.canlet.2018.06.025

Figure 4. Lung metastases are unaffected by in vivo Rapamycin/ICI treatment.

Figure 4

(A) Representative images of lung metastases in mice treated with Rapamycin/ICI for 14 days. Scale bars, 100 μm. (B) Number of lung lesions per cm2 tissue. (C) Size of lung lesions. (N=4 mice; mean ± S.D.). ** p<0.01 by unpaired Student’s t-test. (D) Apoptosis, indicated by immunohistochemical staining for cleaved caspase-3, in primary tumors from untreated hosts, primary tumors from hosts treated with Rapamycin/ICI for 2 days, and lung metastases from hosts treated with Rapamycin/ICI for 14 days. Scale bars, 100 μm. (Positive cells quantified in Supplementary Figure 3A). (E) pS6RPS235/236 staining in primary tumors from untreated and ICI/Rapamycin-treated (2 day) hosts and lung metastases from untreated hosts. Scale bars, 100 μm. (Positive cells quantified in Supplementary Figure 3B). (F) Proliferation in primary tumors and lung metastases from untreated hosts, indicated by Ki-67 staining. Scale bars, 100 μm. (G) Quantification of Ki-67 positive cells from (F) (N= 4 mice; Mean ± S.D.). Different letters denote significant differences (one-way ANOVA followed by Tukey’s multiple comparison tests), p<0.05.