FIG 3.

Differential localization of nuclear and cytoplasmic LANA in migrating Vero cells. (A and B) Vero cells plated in a 16-mm “spot” culture were infected with KSHV. At 24 hpi, the cultures were fixed and stained for LANA (green) and nuclei (blue). (A) Sparse edge of the spot culture, delineated with a dotted white line. The arrows indicate cells with diffuse LANA fluorescence in the cytoplasm located at the edge of the cell culture where the cells were expanding into the open area of the plate. The arrowheads indicate adjacent cells with punctate LANA fluorescence in the nucleus. (B) Center of the confluent spot culture. The arrowheads indicate examples of cells with punctate nuclear LANA fluorescence. (C) Confluent Vero cell cultures were scratch-wounded to stimulate cell migration and proliferation. After a 24-h migration period, the cultures were infected with KSHV. At 24 hpi, the cultures were fixed and stained as described above. The white line marks the edge of the confluent culture following wounding and represents the start point of cell migration. The arrows indicate cells in the migration area with diffuse LANA fluorescence in the cytoplasm (marked with a yellow dot in the nucleus). Adjacent cells and cells in the confluent area had punctate LANA fluorescence in the nucleus. (D) The positions of cells with cytoplasmic LANA relative to the wound edge are shown without the blue and green channels. Scale bars: A and B, 20 μm; C and D, 100 μm.