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