Cover: Three-hour uninfected cells (A–D) and infected cells (E–H) were stained with phalloidin/Alexa 488–labeled (green) and monoclonal anti–ß-tubulin antibody and secondary antibodies Alexa 594–labeled anti-mouse IgG (red) under the same conditions. (A and E) Containing only ß-tubulin label. (B and F) Containing only F-actin label, corresponding fluorescence images to C and G. (C and D) Uninfected cells containing both labels. F-actin (green) is typically present at the basal portion of the cell, around the cell periphery, as a dense lattice of short branching irregular filaments organized into bundles, stress fibers (C), and as punctuate deposits or very short filaments (D). ß-Tubulin (red) is distributed diffusely throughout the cytoplasm, which form a relatively dense network of microtubules in irregularly shaped regions. ß-Tubulin is perinuclearly concentrated. (G and H) Infected cells containing both labels. ß-Tubulin and F-actin are colocalized (yellow) and distributed to a perinuclear local aggregate. F-actin was localized deeper within the cell cytoplasm, ß-tubulin is closer to the plasma membrane. Three-hour infected cells (I and J) and uninfected cells (K and L) were incubated with rhodamine-PE–labeled lipid (red) in presence of unlabeled SP-A, fixed, permeabilized, and stained with monoclonal anti–ß–tubulin antibody and secondary antibodies Alexa 488–labeled anti-mouse IgG (green). (I and J) Internalized lipid (red) in infected cells colocalize with the ß-tubulin marker (green). (K and L) In uninfected cells lipid label (red) and ß-tubulin label (green) are in part separately distributed. Pseudo-color blue were used to highlight the contours of the cell (D, H, I–L). Bar,10 µm. For more information see article by Wissel and colleagues beginning on page 303.
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