Fluorescent protein tagging of adenoviral proteins pV and pIX reveals ‘late virion accumulation compartment’
Fig 5
pV forms ring-like structures within the nucleus before congregating into the LVAC.
(A) Infection progression of HAdV5 pV-mCherry infected cell. (B) Infection progression of HAdV5 pIX-mCherry infected cell. The cells were selected for imaging in live-cell spinning-disk confocal fluorescence microscopy at 24 hpi. The initial nuclear boundaries for the HAdV5 pV-mCherry or pIX-mCherry infected cell are visualized as a dashed line. The cells were imaged as z-stack to be able to adjust for z-movement of infected cells. The image histograms along infection progression have been modified to adjust for changes in protein intensity. (C) Co-stain of pV showing the ring-like phenotype and DBP replication centers in HAdV5 wt infection. A549 cells were infected with HAdV5 wt, fixed at 24 hpi, stained with Hoechst 33342 (Hoechst), and immunostained against pV (anti-pV) and DBP (anti-DBP). Cells were imaged by confocal laser-scanning fluorescence microscopy. The signal overlap is represented in color (merge). A DBP replication ring is enlarged (zoom) with colored corners indicating the channel color. Scalebars indicate 10 μm.