Figure 7. Slug enhances prometastatic phenotypes of PCa cells in vivo.

(A) Fertilized special pathogen–free eggs were incubated for 10 days at 38°C in a rotary humidified incubator. After 10 days of incubation, small holes were drilled over the air sac and near the allantoic vein. 2 × 10⁶ cultured human prostate adenocarcinoma cells (stable LNCaP-vec and LNCaP-Slug cell lines) were implanted onto the membrane in each egg. After sealing the windows, the eggs were incubated in a stationary incubator for 7 days and the embryos were sacrificed after 17 total days of incubation. The embryonic livers and lungs were harvested and analyzed for the presence of tumor cells using quantitative human Alu-specific PCR. (B) LN-SLUG or control cells (150,000 cells/mouse) were injected via the tail vein, after which whole organs were harvested, sectioned completely through; total number of fluorescent cells are reported/organ (left) and representative images shown (right). (C) C42-SLUG-GFP or C42-vec-RFP cells were injected into the left ventricle of nude mice along with fluorescent beads (used as a marker of proper injection). One hour after injection, organs were harvested, whole tissues were sectioned through, and total number of cells counted under a fluorescent microscope. Cells homing to the lung and liver are shown (right and quantified left). Scale bars: 200 μm. Error bars represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.