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. 2021 Jul 22;220(10):e202104093. doi: 10.1083/jcb.202104093

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© 2021 Frazer et al.

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Figure 4. — Prolonged docking of the centrosome to the synapse promotes granule delivery to the synapse. GzmB-TdTomato (red) OTI CTLs expressing Lifeact-EGFP (green) and BFP-PACT (white sphere) interacting with EL4 (blue), pulsed with N4, T4, or G4 peptides. (a–c) Representative maximum-intensity projection time series of OTI CTLs encountering APL pulsed targets. Scale bars = 5 µm; time min:s after contact with target. (d–f) Normalized heatmap of granule-to-centrosome distances from segmented CTL target interactions shown in Fig. 3 b. Color scale represents the density of granules relative to the maximum density on each plot. Total granule-to-centrosome distances measured, n = 15,192 (d), n = 20,343 (e), and n = 16,443 (f). (g–l) R was used to filter granules for concomitant centrosome docking and granule delivery (<0.5 µm of the synapse) and plot them as red spots on a blue trace showing centrosome distance from the synapse. Two representative cells are shown per APL, from a total of N4 = 10, T4 = 9, and G4 = 10.