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. 2014 Jan 13;111(4):1521–1526. doi: 10.1073/pnas.1320638111

Fig. 3.

Fig. 3.

GRK3 induced angiogenesis in vitro and in vivo. (A) GFP- and GRK3-expressing PC3 cells proliferated at similar rates in vitro. These two cell lines were cultured in media with various concentrations of serum for 3 d. Shown on the y axis are growth rates (expressed in fold change) at different concentrations of serum as normalized to 0% serum (x axis). (B) GFP and GRK3-expressing PC3 cells migrated or invaded at similar rates in vitro. Migration or invasion capabilities of these two cell lines were measured by using Transwell migration or Matrigel invasion assays. Error bars refer to SEM. (C) GRK3-expressing PC3 cells were able to induce migration of human endothelial cell seeded in the upper chamber in Transwell assay. Shown on the y axis are numbers of migrated endothelial cells per high-power field. Error bars refer to SEM. (D) MVD is lower in PC3-GFP (Left) than in PC3-GRK3 primary tumor tissues (Middle). Vessels are stained in red with mouse CD34 (green arrows), and dark blue nuclei stains with mouse Ki-67 indicates proliferating endothelial cells (black arrows). Lung micrometastasis (Right) shows ingrowth of proliferating microvessels by coexpression of the endothelial marker CD34 (red) and endothelial cell proliferation by Ki-67 (blue) (magnification of 400×). (E) Representative CD31 staining in red and DAPI staining in blue for prostate tumors from LN4 cells expressing scrambled shRNA or GRK3-shRNA-1.