Figure 7.

Fitness of WDV insertion variants

(A) Quantification of packaging titer of AAV-DJ and 10 different WDV insertion variants via qPCR. Data are means ± SEM. (B) Infection fitness of the WDV insertion variants in (A) quantified by measuring the percentage of tdTomato-positive cells 48 h post transduction at the MOIs of 1 × 10², 1 × 10³, 5 × 10³ and 4 × 10⁴ vg/cell. Data are mean ± SEM. (C) Zoom to the 2-fold and 5-fold axes (outlined) of the capsid surface. Positions of N664 and K708 are shown as red spheres. (D) Quantification of packaging titers of AAV-DJ and WDV insertion variants N664 and K708. Data are mean ± SEM. (E) Infection fitness of the WDV insertion variants N644 and K708 quantified by measuring the percentage of tdTomato-positive cells 48 h post transduction at an MOI of 1 × 10⁴ vg/cell. If indicated, cells were co-expressing GFP-GPI and/or an ssDNA-anti-GFP antibody was added. Data shown as boxplots. Lower and upper hinges of boxes indicate 25^th and 75^th percentile, respectively. Mean is indicated by a horizontal bar in each box. Whiskers extend 1.5 × IQR. Two-way mixed measures ANOVA was used to test the significance of variance between means (AAV-DJ, N664, K708) for conditions with and without GFP present. Presence of the monoclonal antibody (mAb) was a significant source of variation when GFP-GPI was expressed (p value 0.0063) but not without (p value 0.514). Significance levels and p values for pairwise comparisons using a Bonferroni correction are shown.