Figure 1. Production of AAV6 via triple transient transfection at low cell densities. Cells were transfected at 1 or 2 million cells per mL, and plasmid DNA was delivered on a volumetric basis (VB - 1 µg/mL) or cell basis (CB - 1 µg/10⁶ cells). A) Cell growth kinetics and viability. Black arrows indicate the time of transfection, and dashed arrows indicate the time of viral harvest. B) Transfection efficiency is described as the percentage of cells expressing the transgene (GFP) 24 hours post-transfection. C) Volumetric (VG/mL) and cell-specific viral yields (VG/cell). Ctrl: transfection at 1 × 10⁶ cells/mL; 2M: at 2 × 10⁶cells/mL. Values represent mean ± standard deviation (n=3). ** P< 0.01, * P < 0.05, by ANOVA followed by Dunnett’s test.