JCS03483 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Additional images showing ultrastructure of Golgi complex following nocodazole treatment of uninfected and poliovirus-infected cells. Arrows indicate mini-stacks of Golgi complex.

• Supplemental Figure S2 -

  Fig. S2. Effect of nocodazole treatment on Golgi dispersion and virus replication complexes in poliovirus-infected cells. The localization of a Golgi marker, GM130, and a poliovirus replication marker, 2C, were monitored by confocal immunofluorescence microscopy. HeLa cells were pretreated with 10 μg/ml nocodazole (+Noc) or with medium without nocodazole (−Noc) for 1.5 hours Cells were then mock-infected (uninfected) or infected with poliovirus (MOI of 10 PFU/ml) in the presence or absence of 10 μg/ml nocodazole. Cells were fixed at 3.5 hours post infection. Cells grown on coverslips were processed for indirect immunofluorescence and confocal microscopy as shown. The Golgi complex was stained with a murine monoclonal antibody directed against GM130 (red) and an affinity-purified polyconal rabbit antibody against viral protein 2C (green).

• Movie 1 -

  Movie 1 (corresponding to Fig. 5A-C). Example of an uninfected HeLa cell expressing GT-GFP. Notice that the Golgi structure remains as one cohesive structure in the juxtanuclear region of the cell.

• Movie 2 -

  Movie 2 (corresponding to Fig. 5D-E). Example of a poliovirus-infected HeLa cell expressing GT-GFP. Notice that the Golgi structure is dispersed into a haze of fluorescence staring approximately at 60 minutes post infection.

• Movie 3 -

  Movie 3 (corresponding to Fig. 5G-I). Example of an uninfected HeLa cell expressing GT-GFP and treated with nocodazole. The Golgi structure is dispersed into characteristic mini-stacks.

• Movie 4 -

  Movie 4 (corresponding to Fig. 5J-L). Example of a poliovirus-infected HeLa cell expressing GT-GFP and treated with nocodazole. The Golgi structure is dispersed into characteristic mini-stacks but GT-GFP fluorescence does not become diffuse, suggesting that nocodazole treatment prevents further fragmentation of the Golgi vesicles and tubules.