Supplementary information

From the following article:

Imaging the biogenesis of individual HIV-1 virions in live cells

Nolwenn Jouvenet, Paul D. Bieniasz & Sanford M. Simon

Nature 454, 236-240(10 July 2008)

doi:10.1038/nature06998

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Supplementary Information

This file contains Supplementary Figures 1-8 with Legends and Supplementary Methods.

Supplementary Movie 1

This file contains Supplementary Movie 1. Cell expressing a 1:1 mixture of Gag and Gag-GFP, illustrating the transition from a diffuse to a punctate Gag distribution, observed over a 40 minute period. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 2

This file contains Supplementary Movie 2. Example of 3 individual slowly appearing Gag puncta from a cell expressing a mixture of Gag and Gag-GFP observed for 40 min. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 3

This file contains Supplementary Movie 3. An example of a rapidly appearing/disappearing Gag punctum in a cell expressing a mixture of Gag and Gag-GFP observed for 10 minutes. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 4

This file contains Supplementary Movie 4. Appearance of Gag puncta in a cell stably expressing CD63-mCherry (red) and transiently expressing Gag and Gag-GFP (green), observed over a 40-minute period. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 5

This file contains Supplementary Movie 5. Examples of rapidly appearing/disappearing Gag puncta in a cell stably expressing CD63-mCherry (red) and transiently expressing Gag and Gag-GFP (green) observed for 10 minutes. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 15 frames/sec.

Supplementary Movie 6

This file contains Supplementary Movie 6. Examples of a rapidly appearing/disappearing Gag puncta in a cell stably expressing DsRed-clathrin-light-chain (red) and transiently expressing Gag and Gag-GFP (green) observed over a 5 minute period. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 15 frames/sec.

Supplementary Movie 7

This file contains Supplementary Movie 7. Example of a slowly appearing Gag punctum in a cell stably expressing CD63-mCherry (red) and transiently expressing Gag and Gag-GFP (green) imaged over 30 minutes. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 8

This file contains Supplementary Movie 8. Example of a slowly appearing Gag punctum in a cell stably expressing DsRed-clathrin-light-chain (red) and transiently expressing Gag and Gag-GFP (green) observed over 15 minutes. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 9

This file contains Supplementary Movie 9. Cells expressing Gag and Gag-GFP imaged for a total of 28 minutes. The top cell is the control, non-bleached cell. The bottom cell was bleached for 2 minutes after a 5-minute imaging period and observed for 20 additional minutes after the bleach. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

Supplementary Movie 10

This file contains Supplementary Movie 10. View of a portion of a cell imaged for 5 minutes, bleached for 2 minutes and then observed for 20 additional minutes, illustrating both VLPs that recover after the bleach and VLPs that do not. The field of observation is 5.5x5.5μm. The movie was acquired at 1 frame every 5 sec, as described in 'Materials and Methods' and displayed at a rate of 30 frames/sec.

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