Fig. 3

Non-specific adhesion of CoA-dye to cells and glass in the labeling reaction. A Left: representative TIRF images of cells incubated with a reaction mix including different CoA-Atto 565 concentrations (20-min reactions, 5 μM Sfp). Each field of view shows a single transfected labeled cell surrounded by dyes non-specifically adsorbed to glass or other non-transfected cells (see the “Methods” section). Scale bar: 5 μm. Right: quantification of spot density for non-specifically adsorbed dyes using different CoA-Atto 565 concentrations in the reaction, at 1 and 5 μM of Sfp (mean ± SEM from 12–38 fields of view analyzed in two independent replicates). Spot density is measured after 20 min of reaction and extensive sample washing (see the “Methods” section). B Non-specific dye adhesion varying Sfp concentration, at different fixed CoA-Atto 565 concentrations (box: mean ± SEM, whiskers: standard deviation, empty circles: averages, horizontal lines: medians, diamonds: individual data from two independent replicates). ***P < 0.001, **P < 0.01, 1-way ANOVA, Bonferroni multiple comparisons; no significant difference if nothing is shown