Top: representative confocal images of SLP-76 clusters at different times after activation (1.5 min, 5 min and 10 min). Bottom: extrapolation of the copy number of SLP-76-HaloTag per cluster using different DOLs (8%, 12% and 20%). In a , DOL 8%: n = 21 cells; DOL 12%: n = 20 cells; and DOL 20%: n = 9 cells. Cluster analysis for 8% DOL 1.5 min: n = 110 clusters; 5 min: n = 268 clusters; and 10 min: n = 72 clusters. For 12% DOL 1.5 min: n = 59 clusters; 5 min: n = 175 clusters; and 10 min: n = 41 clusters. For 20% DOL 1.5 min: n = 65 clusters; 5 min: n = 237 clusters; and 10 min: n = 41 clusters. Error bars of data points represent DOL: mean ± s.e.m.; cluster analysis: geometric mean of log-normal fit ± s.e.m.; black line: best fit of linear regression; shaded region: 95% confidence interval of linear fit.
