General Instructions
For best detail, please turn up the brightness and contrast on your computer
monitor to the highest settings. Also, make sure your monitor is set to the
maximum resolution in your Mac OS or Windows 95/98/NT system control panel
("millions of colors" on Mac systems, and "32-bit true color" on PC/Windows
95 systems).
Movies show translocation of PKCtheta-GFP
expressed in a T cell clone in response to stimulation with B cells loaded
with specific antigen. The T cells are the bright cells, and the B cells are
the dark "ghost" images.
5 minute
initial translocation movie
Live cell fluorescent imaging shows T
cells (bright cells) interacting with antigen-loaded B cells (dark
"ghosts"). In the center of the image, a T cell forms a conjugate
with a B cell, and T cell PKCtheta-GFP rapidly relocalizes to the T
cell/B cell interface. This movie spans approximately 5 minutes, and
each frame is a 1 second exposure
Multiple
interactions and multiple translocations
This movie spans approximately 20
minutes (each frame is a 10 second exposure), and shows a T cell
making contacts with two different B cells. At both contact points,
T cell PKCtheta-GFP rapidly redistributes to the T cell/B cell
interface. Multiple PKCtheta-GFP translocation events can thus occur
simultaneously in a single T cell.
This movie spans approximately 10
minutes (each frame is a 1 second exposure) and shows a T cell
approaching and contacting an antigen-loaded B cell. PKCtheta-GFP
translocation occurs within 10 seconds following contact with the B
cell. When a 100-fold lower antigen concentration is used,
translocation is delayed for 2-5 min following initial contact (not
shown).
