Analysis of the roles of microtubules in the formation of virus replication centers in the perinuclear region.

<p>U251 cells were infected or uninfected with EV71 for 22°C in the presence or absence of 3 μM Nocodazol for another 2 h. In one treatment, the Nocodazol added was removed (+N removed) by washing with DMEM and then cultured for another 1 h. Cells in all treatments were then fixed for immunofluorescence studies using antibody raised against EV71 or BrUTP to detect the virus replication centers or rhodamine-conjugated DNA probe complemented with EV71 VP1 gene to detect the EV71 genome. (EV71 gen). Microtubules were labeled with antibody specific for alpha-tubulin (tubulin). In the figure, the panels from top to bottom are: EV71 infected cells (+EV71), uninfected control (−EV71), infected cells treated with Nocodasol (+EV71 + Noc), and infected cells with the Nocodasol removed (+EV71 + Noc released). CM: Cell morphology. Bar: 20 μm.