The effect of siRNA mediated silencing of PIK3CA on protein expression, cell viability, migration and anchorage independent growth of FK16A cells. (A) Protein expression of PIK3CA, p-PKB/AKT, total PKB/AKT and actin in FK16A cells (p122) transfected with non-targeting control siRNAs and siRNAs targeting PIK3CA. (B) Cell viability as measured 4 days post-treatment using an MTT assay. Anchorage independent (p119) FK16A cells were transfected with non-targeting control siRNAs, siRNAs targeting PIK3CA. Silencing PLK1 served as a positive control. Appropriate controls were set to 100%. * Indicates a statistical significant difference. (C) Representative pictures of migration assay performed with anchorage independent FK16A cells (p125). Upper panel: cells transfected with non targeting siRNA pool, lower panel: cells transfected with siRNA pool against PIK3CA. Pictures were taken immediately after scratch induction and 48 hours post-induction. (D) Late passage FK16A cells (p119) were seeded in semi-solid agarose following transfection with non-targeting control siRNAs or siRNAs targeting PIK3CA. After 3 weeks in culture colonies were quantified relative to the appropriate controls, set to 100%. * Indicates a statistical significant difference. P: passage.