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. 2017 Jul 11;8(34):57174–57186. doi: 10.18632/oncotarget.19170

Figure 3. Effect of miR-223-3p on tumor biology.

Figure 3

(A) At day 14, the mice were sacrificed and the tumors were extracted for measurement. Despite the effect of miR-223-3p on cell proliferation, we observed no significant difference between the two experimental groups (n = 17). (B) Cell proliferation (evaluated by Ki67 staining) within tumors from mice injected with CAL27 miR-223-3p increased. Representative results are shown in the panels on the left and the number of proliferating cells (nuclear Ki67 staining) per tumor was determined as indicated in the Materials and Methods section. (C) Similarly, the number of apoptotic cells (evaluated by Cleaved Caspase-3 staining) increased in tumors from mice injected with CAL27 miR-223-3p. Representative pictures are shown on the left. Quantification of the number of apoptotic cells, on the right. (D) Immunoblot analysis of ERK2, AKT, STAT3 and their active phosphorylated forms in tumors from CAL27 and CAL27 miR-223-3p mice. Results showed that STAT3 protein is downregulated in cells that express miR-223-3p. (E) The hypoxic status of the tumors from both CAL27 and CAL27 miR-223-3p injected mice is comparable. CA9 staining was used as a read-out of hypoxia. Representative pictures are shown on the left. Quantification of the number of hypoxic cells, on the right. Data, shown as arbitrary units, are representative of 5 mice per group (mean ± sem).*P ≤ 0.05. Bar = 100 μm.