Figure 3

Phenotypic effects of siRNA-mediated knockdown of ANXA10. (A) ANXA10 knockdown in the bladder cancer cell line SW780 was validated by RT–pPCR (left) and western blotting analysis (right) at 48 h post-transfection. Cells were transfected with 5, 10, 50 nM of ANXA10 (ANXA10), 10 nM Control siRNA (Control), no siRNA with transfection reagent (Mock+Lipo), or no siRNA or transfection reagent (Mock). A concentration of 10 nM siRNA was used for western blotting (right). Both experiments were performed in duplicate. (B) The SW780 bladder cancer cell line was transfected with 50 nM siRNA against either ANXA10 (purple) and 50 nM control siRNA (red). The cell index (CI) describes the degree of cell confluence. (C) Wound-healing experiments were performed by wounding confluent SW780 cells 96 h post-transfection with either anti-ANXA10 siRNA or control siRNA (left). ANXA10 and S100A4 expression were measured by RT–qPCR and normalised to Ubiquitin B expression at 72 and 96 h post-transfection (right). Control samples were normalised to 1. ANXA10 knockdown was validated by western blotting analysis at 72 and 96 h post-transfection.