Supporting Movie 2

Movie 2.

Live cell imaging analysis of PKCq -CFP and Bcl10-YFP redistribution in response to antigen stimulation: PKCq and Bcl10 become colocalized at the c-SMAC. D10 T cells expressing PKCq -CFP and Bcl10-YFP were incubated with antigen-loaded APC. T cell/APC interaction and PKCq translocation had already commenced before initiation of data acquisition. This movie shows pseudocolor intensity representations of PKCq -CFP (Left) and Bcl10-YFP fluorescence (Right), where the color spectrum represents fluorescence intensity (red, strongest fluorescence; dark blue, weakest fluorescence), and it contains 40 z-series projection images acquired at 1-min intervals. Note the colocalization between enriched PKCq and enriched Bcl10 at the T cell/APC interface (c-SMAC). The persistent PKCq enrichment at the c-SMAC differs from other data in this report, in which PKCq translocation is of a more transient nature. However, as observed in this movie and in additional data, this phenomenon appears to follow the activation-associated "rounding" of the cell, when it is accompanied by a reorganization of the T cell/APC contact from a broad contact region to a very small, focused region of contact (as occurs in frames 7-12 of this movie).