Figure 5

Activity of PEI/ASO-116 in SK-Mel-29 cells and effect on tumor xenografts in vivo. (A) Semiquantitative RT-PCR analysis of endogenous p73 isoforms in melanoma cells. (B) Quantification of ΔEx2/3p73 transcript levels in melanoma cells transfected with 250 nM PEI/ASO-116 by real-time PCR at indicated time points. Expression was normalized to RPS9. Fold changes are relative to ASO-nsc. Data represent the mean ± S.D. of three independent experiments. (C) BrdU incorporation of cells transfected with PEI/ASO-116- or ASO-sc on day 1 and 3 was measured by ELISA. Significant differences are labeled with asterisks (* p < 0.01; ** p < 0.005). (D) In vivo imaging of PEI/ASO complex 1, 8, and 24 h after intratumoral injection. The fluorescence image (pseudocolor) was overlaid on the photographic image. Intensity of fluorescent signal from Oregon green 488 labeled ASO is shown in top and side view. (E) Relative tumor volumes (RTV) of SK-Mel-29 xenografts injected with PEI/ASO (sc and 116) or MNB/PEI/ASO (M sc and M 116) at a daily interval (p < 0.001). Significant differences between M 116 and 116 are labeled with asterisks (*p < 0.05). (F) ΔEx2/3p73 and TAp73 transcript levels in tumor tissues after 6 day treatment shown in E were quantitated by qRT-PCR. Expression was normalized to RPS9. Fold changes are relative to scrambled controls (set as 1). Bars indicate mean values ± S.D. of n = 4 for each treatment.