Fig. 4: CCDC719-13 encodes a protein that exerts its function extracellularly.
A Representative images of colony formation assays in PC3 and DU145 cells treated with supernatant collected from PCa cells transfected with Vector or CCDC719-13. Colonies were stained with crystal violet. B Quantification of the number of colonies formed by PC3 and DU145 cells. Data represent the mean ± SD from three independent experiments. C Relative cell viability of PC3 and DU145 cells treated with supernatant from PCa cells transfected with Vector or CCDC719-13 over a 6-day period, assessed using the CCK-8 assay. Data represent the mean ± SD from three independent experiments. D Flow cytometry analysis of apoptosis in PC3 and DU145 cells treated with supernatant from PCa cells transfected with Vector or CCDC719-13. E Quantification of apoptotic cells in PC3 and DU145 cells. F, G ELISA measuring CCDC719-13 levels in PC3 and DU145 cells transfected with Vector, CCDC719-13, or supernatant from cells treated with Brefeldin A (BFA) to block protein secretion. H, I Immunofluorescence analysis of CCDC719-13 expression in PC3 and DU145 cells treated with supernatant from PCa cells transfected with Vector or CCDC719-13. Cells were stained with anti-Flag antibody (red) and DAPI (blue) to visualize nuclei. J, K Relative cell viability of PC3 and DU145 cells treated with supernatant from PCa cells transfected with Vector, CCDC719-13, or supernatant from cells treated with BFA, over a 6-day period, assessed using the CCK-8 assay. L Representative images of colony formation assays in PC3 and DU145 cells treated with supernatant from PCa cells transfected with Vector, CCDC719-13, or supernatant from cells treated with BFA. Colonies were stained with crystal violet. M, N Quantification of the number of colonies formed by PC3 and DU145 cells. Data represent the mean ± SD from three independent experiments. Quantification of cell death in PC3 (O) and DU145 (P) cells treated with conditioned medium derived from prostate cancer cells transfected with Vector or CCDC719-13, with or without Brefeldin A (BFA) pretreatment. Cell death was measured by 7-AAD staining followed by flow cytometry.
