FIG. 6.

Morphological distribution of M1 and HA in influenza virus-infected MDBK cells by confocal microscopy. MDBK cells (4 × 10⁵) were grown on chamber slides and synchronously infected with WSN virus at an MOI of 10 at 4°C. Cells were then washed and incubated at 37°C. Monensin at a 10 μM final concentration was added to some cells at 2 hpi (G to L), and the cells were incubated for another 5 h at 37°C. At 7 hpi, all virus-infected cells were fixed with ice-cold acetone, incubated with a mixture of anti-M1 rabbit polyclonal and anti-HA mouse monoclonal antibodies, and stained with anti-rabbit IgG (green) and anti-mouse IgG (red). The stained cells were examined by confocal microscopy as described in Materials and Methods. (A to C) Mock-infected cells without monensin; (D to F) virus-infected cells without monensin treatment; (G to I) mock-infected cells with monensin; (J to L) virus-infected cells with monensin. Image analysis was done as follows: panels A, D, G, and J for M1 (green); panels B, E, H, and K for HA (red); and panels C, F, I, and L superimposed for both HA and M1 (original magnification, ×1,000).