Figure 5.

Transduction of all tumor layers by AAV2 vector in TGFα/c-myc HCC mice. Tumor-bearing animals were i.v.-injected with scAAV2.CMV.eGFP. Vector dose: 1 × 10¹¹ particles per animal. Four days post-administration, HCC lesions were isolated, and transgene (eGFP) expression was analyzed from histological images. Enhanced GFP-positive cells in HCC were visualized by immunostaining. Enhanced GFP-positive cells per total tumor cells were quantified from random images of 30% of the tumor area of each lesion. Random images were assigned to distinct tumor layers defined as the border, the first layer, the second layer, and the core. (A) eGFP-positive cells in tumors comprising the border; n = 12. (B) eGFP-positive cells in tumors comprising the border and first layer; n = 12. (C) eGFP-positive cells of tumor lesions comprising the border, first and second layer; n = 5. (D) eGFP-positive cells of tumor lesions, which comprise all defined layers; n = 14. Data points are labeled by sample number of analyzed tumor, 1 to 43. Data are shown in log₁₀-scale as the average radiance with the grand mean within each cohort. Statistics: Cohort sizes of (B,D) account for the Shapiro–Wilk normality test (p < 0.0001); Shapiro–Wilk normality test with log₁₀-transformed data ((B) p = 0.0147; (D) p = 0.0427). (A) Mann–Whitney test (ns); (B) ordinary one-way ANOVA with log₁₀-transformed data (ns) and Tukey’s multiple comparisons test (ns), (C,D) Kruskal–Wallis test (ns), and Dunn’s multiple comparisons test (ns).