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.

Indirect immunofluorescence confocal microscopy was used to investigate the localization and possible abundance of the mitochondrial proteins in HRSV-infected cells. The NDUFB10 localization validation indicated that the subcellular localization of this protein was altered in HRSV-infected cells when compared with mock-infected cells, and increased fluorescence intensity was also observed in the former (Fig. 7A) . The fluorescence data for the subcellular localization of rep-resentative TOM complex members (Tom20 and Tom22) indicated that the subcellular localization of these proteins also differed between HRSV-infected cells and mock-infected cells with localization observed in a distinct region of the cytoplasm. There also appeared to be less fluorescence from Tom20 and Tom22 in HRSV-infected cells (Fig. 7 , B and C, respectively). In addition, the data indicated that in HRSVinfected cells Tom20 was possibly present in the nucleus or at least closely associated with this structure. This was confirmed in the Z-stack image of this cell (supplemental Fig. 1) . The relative fluorescence of VDAC1 in HRSV-infected cells was greater than in mock-infected cells. The subcellular localization was also altered with increased punctate staining observed (Fig. 8A) . The subcellular localization of PHB in HRSV-infected cells appeared to be localized with viral complexes and with greater fluorescence intensity in these regions than in mock-infected cells (Fig. 8B) .