Promising Noninvasive Cellular Phenotype in Prostate Cancer Cells Knockdown of Matrix Metalloproteinase 9
Figure 2
Analysis of surface expression of CD44v6 in indicated PC3 cell lines. (a) and (b), Surface expression of CD44v6 in PC3 cells knockdown of MMP9 (PC3/Si) is compared with control cell lines (PC3, PC3/V, and PC3/Sc) by FACs analysis (a) and (b). A representative histogram demonstrating the surface levels of CD44v6 in various PC3 cell lines is shown. In a typical experiments about 4–6 wells in 24 wells were used for each cell line. Statistical analysis of the mean fluorescence intensity (mean ± SEM; ) is shown as a graph (b); * versus control cell lines. (c) and (d) Immunoblotting analysis of surface expression of CD44v6 and Zip1 proteins in indicated PC3 cell lines. Equal amount of lysate proteins (100 μg) were immunoprecipitated with an antibody to CD44v6 (c) and Zip1 (d) and pulled down with streptavidin agarose. The blot was probed with a CD44v6 antibody followed by a Zip1 antibody after membrane stripping. A shorter exposure blot for PC3/Si is shown in lane 5. (e) Deglycosylation of CD44v6 with TFMSA. Total lysates (500 μg protein) made from PC3/Si cells were treated with (+) and without (−) TFMSA. Immunoblotting analysis demonstrates the deglycosylated (lane 1) and non-deglycosylated (lane 2) CD44v6 (e). The results shown are representative of three different experiments.
