Microscopy
HEK-293SF cells were seeded at low confluence on a 35mm plate with
coverslip (MatTek). 24h post seeding, the cells were transfected for the
production of AAV2-GFP. 72h post seeding, the media was removed, and the
cells fixed and stained as previously described (Kerviel et al., 2016).
AAV capsid monomers were detected with anti-VP1/VP2/VP1 (Progen)
labelled with AlexaFluor 700 (Invitrogen), hAd5 E2A by anti-E2A labelled
with AlexaFluor 350 (Invitrogen). Anti-E2A was a gift from Arnold J.
Levine (Reich, Sarnow, Duprey, & Levine, 1983). Antibodies were
incubated at room temperature for 1 hour. Cells were then imaged using
an Olympus IX-83 confocal microscope. The images were analyzed using
FIJI.