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. 2014 Jan 16;5(5):1265–1278. doi: 10.18632/oncotarget.1711

Figure 3. Evaluation of the anti-proliferative effects of EGFR endocytosis inhibitors.

Figure 3

(A) Inhibition of of EGFR endocytosis was induced by treatment with dynasore and dynole 34-2, or a combined treatment with gefitinib in H1703 cells. The altered internalization of EGFR endocytosis was measured by immunofluorescence staining with a specific antibody against EGFR. The immunofluorescence images were obtained using a fluorescence microscope at a magnification of 400×. (B) Effects of EGFR endocytosis inhibitors alone or combination with gefitinib on cell viability were analyzed by the MTT assay. The cells were treated with dynasore (100 μM) or dynole 34-2 (10 μM) or in combination with gefitinib (10 μM) for 48 h. Each bar represents mean values acquired from three experiments with standard error (SE). * p < 0.05. (C) The rate of apoptosis was investigated by Annexin V-FITC staining. The percentages of Annexin V-positive cells were examined by flow cytometry under the same conditions as the MTT assay. Each bar represents mean values acquired from three experiments with SE. * p < 0.05. (D) Changed expression levels in apoptosis related proteins were evaluated after treatment with EGFR endocytosis inhibitors and gefitinib using Western blot analysis. Cell lysates were blotted for the indicated proteins with the appropriate antibodies after cell harvest.