Fig 1: PKN1 and PKN2 siRNAs increase expression of AXIN2 in melanoma cell lines. The indicated cell lines were transiently transfected with siRNA oligonucleotides targeting PKN1 and PKN2 (pan PKN) or targeting nonsense sequence (CTRL). Forty-eight hours after transfection the cells were stimulated overnight with medium conditioned with WNT3A or with vehicle (left panel, dose curve; right panel, single dose). The effects of WNT3A were assayed using quantitative PCR. cDNA was amplified using primers targeting GAPDH and the Wnt/β-catenin target gene AXIN2. The number of copies of AXIN2 cDNA was averaged, normalized to 1 × 105 copies of GAPDH amplified from the same sample, and plotted. The error bars represent S.D., and the data in all plots are representative of at least two independent experiments.