Quantitative Proteomic Analysis of A549 Cells Infected with Human Respiratory Syncytial Virus

Munday, Diane C.; Emmott, Edward; Surtees, Rebecca; Lardeau, Charles-Hugues; Wu, Weining; Duprex, W. Paul; Dove, Brian K.; Barr, John N.; Hiscox, Julian A.

Using Ingenuity Pathways Analysis, these proteins were grouped into functional classes and used to potentially map Nuclei are stained blue with Hoechst. All mitochondria are stained red with Mito-Tracker (A), healthy cells are also stained green with calcein AM (B), which is concentrated in the mitochondria. Merged images are also presented (C). Positive control cells were treated with ionomycin to allow the entry of excess calcium into the cells to trigger mitochondrial pore activation, which leads to the release of calcein AM and therefore the loss of green florescence. A large proportion of mitochondria in HRSV-infected cells were stained red and only weakly green, indicating that HRSV infection affected mitochondrial transition pore activity. Scale bars, 20 m. Note that indirect immunofluorescence confocal microscopy was used to demonstrate that cells were infected with HRSV, and these data are presented in supplemental Fig. 2. the nuclear and cytoplasmic proteomes of A549 cells (Fig. 2) . The nuclear proteome of A549 cells has been investigated previously using 2DE and HPLC, highlighting the extensive use of A549 cells in respiratory disease research (33) . Therefore, our current study complements and expands this analysis and adds further data on cytoplasmic proteins.