Fig. 2.

Microcluster formation precedes VAMP7 recruitment as visualized by TIRF microscopy. a Jurkat cells transfected with LAT-RFP and GFP-VAMP7 were dropped onto stimulatory coverslips and imaged by TIRF microscopy. b Intensity profile of LAT-RFP and GFP-VAMP7 over time. c Time of appearance of LAT-RFP or GFP-VAMP7 signal in TIRF field (n = 12 cells, three independent experiments). d Jurkat cells transfected with Grb2-apple and emerald-VAMP7 were dropped onto stimulatory coverslips and imaged by TIRF microscopy (n = 10 cells, three independent experiments). a, d Top panels show LAT and Grb2, respectively, and arrows point to early microclusters; bottom panels show VAMP7, and arrowheads point to vesicles touching down. Scale bars = 2 μm