Video 5. Lifetime of LOV2 and Zap70 after optogenetic signal termination.

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Example cell from time course of bound LOV2 (left) and recruited Zap70 (right) loss after inactivation of antigen signaling with strong blue light. Jurkat cells expressing zdk-CAR and Zap70 biosensor were activated on LOV2 and ICAM-1 functionalized supported lipid bilayers (SLBs) for 3 min prior to the start of the video. Cells are illuminated with strong blue light at the start of the video, and the lifetime of bound LOV2 and recruited Zap70 are tracked using the ImageJ TrackMate plugin (magenta circles, MM:SS timestamp top left). Only tracks originating before blue-light illumination (start of the video) were included to filter out spurious tracks later in the time course (scale bar = 5μm).