Figure 4

Inhibition of P-cadherin in HOXA9-overexpressing EOC cells abrogates the stimulatory effects of HOXA9 on cell aggregation, survival, implantation and migration. (A) Western blot analysis of P-cadherin levels in vector-control and HOXA9-transfected SKOV3-Par lines, and in HOXA9-transfected SKOV3-Par lines that stably expressed non-targeting shRNA and shRNAs targeting two different sites within the CDH3 gene (shPCAD-A, shPCAD-B). (B ,C) Cells of SKOV3-Par lines were incubated as suspension cultures in polyHEMA-coated plates for 3 days. (B) Cell morphology viewed by phase-contrast light microscopy. Bar, 50 μm. (C) Evaluation of cell death by flow cytometric analysis of 7AAD staining. (D) Evaluation of cell death by ELISA in suspension cultures of SKOV3-Par cells, attachment of SKOV3-Par cells to confluent mesothelial cell monolayers, and migration of SKOV3-Par cells were assayed as described in Figures 2D, E and G, respectively. Shown in D are mean + sd values of three independent assays.