Fig. 7.

H84T restricts influenza virus to the late endosomal/lysosomal compartment. (A) Representative immunofluorescent micrographs of M1 (green) and LAMP1 antigen staining (red) and nuclei (blue). Following 1 h pretreatment with H84T (10 μM), MDCK cells were incubated with A/WSN/1933 (H1N1) at 4 °C for 1 h to allow only for attachment, excess virus was removed, treatment-containing medium was replaced, and the cells were incubated at 37 °C for an additional 3 h. The second and fourth rows are micrographs of one or two cells from the first and third rows, respectively, indicated by the arrows. Data are representative of five independent experiments. Scale bars indicate 50 μm in the first and third rows and 10 μm in the second and fourth rows. (B) Quantitation of the percent of M1⁺ punctae that are also LAMP1⁺ (percent overlap). Statistical analyses were performed by t test. ****P < 0.0001, as compared to the infected, untreated group. Error bars represent the SEM.