Journal: Scientific Reports

Article Title: Functional roles of the membrane-associated AAV protein MAAP

doi: 10.1038/s41598-021-01220-7

Figure Lengend Snippet: MAAP sequence. MAAP is encoded in the wt-AAV2 genome (GenBank Sequence ID: AF043303.1) at nucleotides 2282-2641. The ORF is located on the same DNA region encoding the VP1/2 unique domains. VP1/2 are encoded on reading frame + 1 and MAAP on reading frame + 2. MAAP translation initiates at a CTG codon (start 1), encoding a leucine which corresponds in position to VP1 proline in position 27. The next potential start codons are MAAP-T13 (ACG) and MAAP-R14 (AGG) (start 2-3). In red are the three basic-amino-acid-rich clusters MAAP2BR1, MAAP2BR2 and MAAP2BR3. Underlined is the sequence of the peptide used to generate MAAP polyclonal antiserum. All studied amino acid substitutions are shown below the sequence.

Article Snippet: MAAP was detected with a specific polyclonal antibody (Ab) (MAAP GAL-KKI, Davids Biotechnologie), viral capsid proteins with a monoclonal antibody (MAb) against VP1, VP2 and VP3 (B1, Progen, #61084), and intact viral capsids with a MAb (A20, Progen, #61055).

Techniques: Sequencing

Journal: Scientific Reports

Article Title: Functional roles of the membrane-associated AAV protein MAAP

doi: 10.1038/s41598-021-01220-7

Figure Lengend Snippet: Localization and intensity of MAAP and AAV-2 capsid proteins in transfected cells. ( A ) Representative confocal images of intracellular localisation of MAAP and capsid proteins at 24 hpt. Cells were co-transfected with wt-AAV2 and MAAP-S33-S39-S47 variant together with Ad helper plasmid, or with recombinant MAAP expression plasmid without Ad helper plasmid. The cells were immunolabelled with polyclonal antibody against MAAP (green) and monoclonal antibody against viral capsid proteins VP1, VP2 and VP3 (red). Blue corresponds to DAPI staining. Scale bars, 3 µm. For analyses, the cells were either mock transfected (n = 19), co-transfected with wt-AAV2 (n = 11), MAAP-S33-S39-S47 (n = 19), MAAP-W105 (n = 20), MAAP-L106 (n = 17), MAAP-L110 (n = 20) together with Ad helper plasmid, or transfected with the recombinant MAAP plasmid (n = 18). ( B ) Quantitative analyses of the total intensities of MAAP (light grey) and capsid proteins (dark grey) at 24 hpt. ( C ) Quantitative analyses of the relative intensity of nuclear (dark grey) in comparison to cytoplasmic capsid proteins (light grey) at 24 hpt. For analysis, the cells were immunolabelled with antibodies against viral capsid protein VP1, VP2 and VP3 (red) and the nuclear boundary segmentation was based on distribution of DAPI-labelled chromatin (blue). The error bars show the SEM.

Article Snippet: MAAP was detected with a specific polyclonal antibody (Ab) (MAAP GAL-KKI, Davids Biotechnologie), viral capsid proteins with a monoclonal antibody (MAb) against VP1, VP2 and VP3 (B1, Progen, #61084), and intact viral capsids with a MAb (A20, Progen, #61055).

Techniques: Transfection, Variant Assay, Plasmid Preparation, Recombinant, Expressing, Staining