FIG 6.
L protein interaction with vRNP distinguishes between DVG-high and FL-high cells. (A) A549 cells infected with SeV LeGFP supplemented with purified DPs (number of hemagglutinating units [HAU] = 20) for 24 h and then subjected to RNA FISH with immunofluorescence for GFP (L, gray) and Hoechst staining (nuclei, blue). Confocal 63×, 1.5× zoom, single-plane images are shown. Images are representative of results from four independent experiments; scale bar = 20 μm. Green boxes indicate representative DVG-high cells; orange boxes indicate representative FL-high cells. (B) Representative cropped DVG-high cell. (C) Representative cropped FL-high cell. (D) Costes’s Pearson’s correlation of colocalization between (-)5′ SeV and SeV NP in individual DVG-high and FL-high cells. (E) Mander’s overlap coefficient of L (GFP) with 5′ SeV in DVG-high and FL-high cells. (F) Mander’s overlap coefficient of 5′ SeV with L (GFP) in DVG-high and FL-high cells. Individual cells pooled from three independent experiments are plotted, with the horizontal dotted line at the mean; error bars represent SD. **** = P < 0.0001 by Mann-Whitney U-test.