Linker for Activation of T Cells, -Associated Protein-70, and Src Homology 2 Domain-Containing Leukocyte Protein-76 are Required for TCR-Induced Microtubule-Organizing Center Polarization

Files in this Data Supplement:

• MTOC Movie 1 (AVI Format, 8,851 KB) - The movement of MTOC is preceded by release of intracellular calcium stores. 1x 10⁶Raji B cells were loaded with superantigen, SED, and added to 5 x 10⁵ JTUB cells that had been loaded with the calcium indicator, FURA-2. The interaction between the JTUB cells and the super antigen-loaded B cells were observed simultaneously as a DIC image (upper left panel), a mid-cell z-section of GFP (lower left panel) as well as a ratio of Fura-2 wavelengths (upper right panel). The ratio of Fura-2 wavelengths was pseudo colored with a color spectrum to reflect the relative concentration of free calcium in the JTUB cells. Cell-cell interactions were imaged every 15 seconds for 15 minutes.
• MTOC Movie 2 (QuickTime Format, 8,856 KB) - See caption above.
• MTOC Movie 3 (AVI Format, 6,965 KB) - See caption above.
• Jurkat Calcium Movie (QuickTime Format, 8,126 KB) - Jurkat T cells make stable conjugates with APCs. Raji B cells loaded with superantigen, SED, were added to 5 x 10⁵ Jurkat cells that had been loaded with the calcium indicator, FURA-2. The interaction between the T cells and the super antigen-loaded B cells were observed simultaneously as a DIC image (left panels) and as a ratio of FURA-2 wavelengths (right panels). The ratio of FURA-2 wavelengths was pseudo colored with a color spectrum to reflect the relative concentrations of free calcium. Cell-cell interactions were imaged every 15 seconds for 15 minutes.
• JCaM2 Calcium Movie (QuickTime Format, 7,532 KB) - JcaM2 T cells make stable conjugates with APCs. Raji B cells loaded with superantigen, SED, were added to 5 x 10⁵ JCaM2 cells that had been loaded with the calcium indicator, FURA-2. The interaction between the T cells and the super antigen-loaded B cells were observed simultaneously as a DIC image (left panels) and as a ratio of FURA-2 wavelengths (right panels). The ratio of FURA-2 wavelengths was pseudo colored with a color spectrum to reflect the relative concentrations of free calcium. Cell-cell interactions were imaged every 15 seconds for 15 minutes.
• J14.V29 Calcium Movie (QuickTime Format, 7,801 KB) - J14.V29 Cells T cells make stable conjugates with APCs. Raji B cells loaded with superantigen, SED, were added to 5 x 10⁵ J14.V29 cells that had been loaded with the calcium indicator, FURA-2. The interaction between the T cells and the super antigen-loaded B cells were observed simultaneously as a DIC image (left panels) and as a ratio of FURA-2 wavelengths (right panels). The ratio of FURA-2 wavelengths was pseudo colored with a color spectrum to reflect the relative concentrations of free calcium. Cell-cell interactions were imaged every 15 seconds for 15 minutes.