Journal: PLoS ONE

Article Title: NGF Is an Essential Survival Factor for Bronchial Epithelial Cells during Respiratory Syncytial Virus Infection

doi: 10.1371/journal.pone.0006444

Figure Lengend Snippet: (A). Dot plots of simultaneous annexin V and 7-aminoactinomycin D (7-AAD) staining of rgRSV-infected bronchial epithelial cells transfected with scrambled (SCR) siRNA (left) or with NGF-specific siRNA (right). After 48 h of transfection, cells were infected with 5 MOI of rgRSV. Apoptosis and necrosis were detected by DNA labeling with 7-AAD-PE and annexin V-Cy3 respectively. Data are expressed as percentage (%) of total cells. (B). Apoptosis vs. necrosis assessed by annexin V and 7-AAD staining followed by flow cytometry. Non-transfected controls include non-infected, rgRSV-infected, and UV-RSV-exposed cells. Data are expressed as the mean±SEM (n = 3 experiments). ** = p <0.01; *** = p <0.001 compared to non-infected controls.

Article Snippet: In separate experiments, cells were treated with both 7-AAD (Enzo Life Sciences, Farmingdale, NY), a fluorescent intercalating dye that allows DNA quantitation, and annexin V-Cy3 for 15 min. After fixation with 1% paraformaldehyde, 500 µl of 1× annexin buffer were added to each sample and fluorescence flow cytometric analyses were performed using a BD Biosciences FACSCaliber instrument.

Techniques: Staining, Infection, Transfection, DNA Labeling, Flow Cytometry