Fig. 3

Influenza A virus infection of mouse respiratory epithelial cells results in greater internalization of the epidermal growth factor receptor (EGFR) in cells derived from males. Mouse tracheal epithelial cell (mTEC) cultures were isolated from male and female mice and either mock-infected or infected with 2009 H1N1 at a high multiplicity of infection (MOI = 10). At 24 hpi, the cultures were fixed and immunostained for the 2009 H1N1 hemagglutinin protein (HA; red). The cultures were then permeabilized and immunostained for the EGFR (green) with a nuclear counterstain (DAPI; blue). a Quantitation of the ratio of internalized to basolateral EGFR fluorescence signal present in panels b and c. Reconstructed Z sections from 0.5 μm optical sections obtained via confocal microscopy showing the localization of HA and EGFR in mock (b) or IAV-infected (c) mTEC cultures derived from male or female mice. The images were obtained on an LSM 700 using a × 63 objective and processed using Volocity and FIJI software. Data represent means ± SEM from two independent experiments, with significant differences represented by asterisks (*). At least eight fields were counted per condition, per experiment