Fig. 2: Ablation of MAAP expression results in a significant delay in the extracellular secretion of wildtype and recombinant AAV8 particles.
From: The membrane associated accessory protein is an adeno-associated viral egress factor
A Schematic of WT AAV8 MAAPΔ mutant. B AAV8 ssCBA-Luc vectors produced with WT Cap or MAAPΔ Cap. Total vector genomes collected from the cells and media and the proportion of virus found in each media harvest or associated with the cells C are shown. Data are presented as mean values ± SD. Significance was determined by two-way ANOVA, with Sidak’s post-test. (Media (D3) c Media (D5) ****p < 0.0001; Cells (D5) ****p < 0.0001, Total ***p = 0.0004). D Schematic of rAAV8 MAAPΔ mutant. E AAV8 ssCBA-Luc vectors produced with recombinant Cap or MAAPΔ Cap. Total vector genomes collected from the cells and media and the proportion of virus found in each media harvest or associated with the cells (F) are shown. Data are presented as mean values ± SD. Significance was determined by two-way ANOVA, with Sidak’s post-test. (Media (D3) **p = 0.0081; Media (D5) **p = 0.0036; Cells (D5) ****p < 0.0001, Total ****p < 0.0001). Recombinant AAV8 and AAV8 MAAPΔ viruses were analyzed from the media and pellet of HEK293 producing cells at days 3 (G) and 5 (H) post transfection. Capsid proteins were analyzed by SDS-PAGE under reducing conditions and probed with a capsid (B1)-specific antibody. Immunoblots are representative images of two independent experiments. I Luciferase assay analyzing transduction of HEK293 cells by AAV8 and AAV8 MAAPΔ mutant virus at MOIs of 10,000 and 50,000 vg/cell. Each bar is a representation of three experiments that are biological replicates.
