Co-localization of NS1 with MAVS in mitochondria during RSV infection.

A549 cells were infected with rA2 or rA2ΔNS2 (0.1 MOI).Twelve hours after infection, cells were stained with the mitochondrial stain CMXRos (red). Cells were then fixed and stained with Alexa 488-conjugated anti-NS1 antibody (green) and Alexa 647-conjugated anti-MAVS antibody (purple). Stained cells were viewed under the Leica TCS SP2 laser scanning confocal microscope (A) Images (400× magnification) show individual staining colors, and merged images of NS1-mitochondria and NS1-MAVS are also shown. (B) The images are a 5× enlargement of a 400× image. The top panel shows the cells infected with rA2 and the bottom panel the cells infected with rA2ΔNS2. The areas marked by white circles show the presence of NS1 in mitochondria and colocalization with MAVS. The first pair of images are merges of NS1 and mitochondria. The second pair show NS1 merged with MAVS and the last pair shows all three. (C). Correlation analysis of the co-localization of NS1 with mitochondria in rA2 infected cells. (D). Correlation analysis of the colocalization of NS1 with mitochondria in rA2ΔNS2 infected cells.Pearson's coefficient for MAVS and NS1 is shown along with Manders' coefficients (M1 & M2), which represent the fraction of the mitochondrial red overlapping with NS1 green and the fraction of NS1 green overlapping with the mitochondrial red, respectively.