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. 2016 Jul 22;8(11):17700–17711. doi: 10.18632/oncotarget.10775

Figure 3. Capz inhibits invasion and promotes apoptosis in prostate cancer cells.

Figure 3

A. An invasion assay was performed using the Roche xCELLigence Real-Time Cell Analyzer (RTCA) DP instrument (Roche Diagnostics GmbH, Germany). DU145 cell invasion activity (4 × 104 cells/well) was measured in a matrigel-coated CIM (cellular invasion/migration)-Plate 16 with the indicated concentrations of Capz. B. DU145 cells were seeded in a matrigel invasion chamber overnight in the absence of a serum, incubated with the indicated concentrations of Capz for 24 h, and then subjected to the invasion assay. Representative photographs of stained cells on the lower side of matrigel membrane from one of the three experiments are shown. C. DU145 cells (1 × 106 cells/well) were treated with 5 μM of Capz for 24 h. The cells were incubated with a FITC-conjugated Annexin V antibody and then analyzed by flow cytometry. D. DU145 cells (1 × 106 cells/well) were seeded into 6 well plates and treated with 5 μM of Capz for 24 h. The cells were fixed and incubated with TUNEL reaction solution and then analyzed by flow cytometry. E. DU145 cells (1 × 106 cells/well) were treated with the indicated concentrations of Capz for 24 h. Whole-cell extracts were prepared; 20 μg portions of those extracts were resolved via 10% SDS-PAGE and probed with pro-caspase-3, cleaved caspase-3, and PARP antibodies. The same blots were stripped and reprobed with β-actin antibody to verify equal protein loading. F. Equal amounts of lysates were analyzed by Western blot using antibodies against Bcl-2, Bcl-xl, and Survivin. The same blots were stripped and reprobed with β-actin antibody to verify equal protein loading. G. Equal amounts of lysates were analyzed by Western blot using antibodies against Cyclin D1 and COX-2. The same blots were stripped and reprobed with β-actin antibody to verify equal protein loading. H. Equal amounts of lysates were analyzed by Western blot using antibodies against VEGF and MMP-9. The same blots were stripped and reprobed with β-actin antibody to verify equal protein loading. The results shown are representative of two independent experiments.