Fig. 7.

Removal of recovered cortical actin does not affect PIP₂ at the synapse. (A, Left) Time-lapse TIRF images of an OT-I CTL expressing Lifeact-mApple (gray scale, Top Row) and Tubby-EGFP (gray scale, Middle Row) or merged channels (Bottom Row) that already had secreted and recovered actin in response to anti-CD3–coated glass surfaces (−0:40) and then were treated at 0:00 with 1 μM LatA. (Right) Kymographs of movies. Fluorescence under a three-pixel-thick region of interest (dashed white line) is displayed over time. Dashed yellow lines indicate the time of treatment addition. (Scale bar: 5 μm.) Time is shown in minutes:seconds. (B) The graph depicts the percentage of cells that cleared reporter (Lifeact, red bars or PIP₂, green bars) at the membrane after treatment with carrier or LatA. Data are shown as mean ± SD from three independent experiments; n = 19 for carrier-treated cells; n = 81 LatA-treated cells.