Figure 6
Reconstitution of P-cadherin in HOXA9-knockdown EOC cells restores the stimulatory effects of HOXA9 on cell survival, implantation and migration in vivo. Female nude mice were inoculated i.p. with equivalent numbers (2 × 106) of cells of control (non-targeting) and HOXA9-knockdown (shA9-B) SKOV3ip lines and HOXA9-knockdown SKOV3ip cells that stably expressed P-cadherin (shA9-B + PCAD). In (A), mice were sacrificed at 3 days following tumor cell inoculation. Floating cells in the peritoneal cavity were collected and stained with 7AAD. Cell death within the gated population of GFP+ tumor cells was evaluated by flow cytometric analysis of 7AAD staining. In (B-D), mice were analyzed at 3 weeks following tumor cell inoculation. (B) Representative examples of GFP-expressing tumor implants (shown in green) attached to the peritoneal cavity wall as viewed by fluorescence microscopy. Bar, 1 mm. Numbers of implants on the peritoneal cavity wall were counted in five random 25 mm2 fields per mouse. (C) Representative examples of HE-stained sections of bowel tissues with mesenteric implants. Bar, 1 mm. Numbers of mesenteric implants were counted in three random 1 cm2 fields per mouse. (D) Representative examples of HE-stained sections of diaphragmatic implants. Bar 100 μm.