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. Author manuscript; available in PMC: 2019 Dec 27.
Published in final edited form as: Cell. 2017 Mar 23;169(1):108–119.e20. doi: 10.1016/j.cell.2017.03.006

Figure 5. Difference in Microcluster Formation by Low- and High-Affinity Ligands with DNA-CARζ.

Figure 5

(A) Formation over time of ligand-receptor clusters (defined as a diffraction limit structure containing ≥ 2 ligands) for individual cells (colors) at 1 ligands/μm2 for the 16-mer and 30 ligands/μm2 for the 13-mer. t = 0 is defined as the point of image acquisition, generally within 1–2 min of adding cells to the SLB.

(B) Distribution of dwell times for ligand-receptor clusters composed of 16-mer (n = 94 from 6 cells; 35 clusters with >100 s dwell times were truncated by the end of image acquisition) and 13-mer ligands (n = 125 from 6 cells; 6 clusters similarly truncated by image acquisition) at 1 ligands/μm2 and 13-mer DNA ligand at 30 ligands/μm2 (n = 203 from 5 cells; 26 clusters truncated by image acquisition).

(C) TIRF images and intensity time series showing the formation of transient receptor-ligand clusters of 13-mer DNA ligand at 1 ligands/μm2. The fluorescence time series were analyzed as described in Figures 3 and S3.

See also Figure S6.