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MBC Video for Molecular Biology of the Cell: Volume 0, Issue 2002 -- Page 02-05-0078

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Supplementary QuickTime Videos

• Supplementary_movie_1.mov

  CD45 co-localizes with lipid rafts upon BCR crosslinking. Bal 17 cells were adhered onto a poly L-lysine coated glass dish and after blocking Fc receptor binding by treating with 2.4G2 , the cells were stained with FITC-conjugated anti-CD45 followed by washing and staining with CTB-TRITC. The dish was fixed on a stage adapter, the microscope focused on the cells and images acquired using the fluorescein and rhodamine filters in the unstimulated state (upper row). Anti-IgM was added to the dish and image acquisition was started simultaneously. Images were collected over a period of 10 min with a time-lapse of 15 sec. Surface planar views of the cells are shown, with CD45 in green, lipid rafts in red and regions of co-localization in yellow indicated by white arrows. Still images corresponding to the movie are shown in Figure 4B.

• Supplementary_movie_2.mov

  Translocation of the Syk tyrosine kinase to the plasma membrane upon BCR crosslinking. Bal 17 cells stably expressing Syk-GFP were stimulated with anti-mouse IgM at room temperature. The cell suspension was placed on a slide and covered with a cover slip. After focusing on cells, image acquisition was started and continued every 10 sec for a period of 10 minutes. All sixty images were saved as a time-lapse movie. Still images corresponding to the movie are shown in Figure 6A.

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