Extended Data Fig. 2: Statistical analysis of cell viability using flow cytometry.
U2OS cells were incubated either with TF-/CF-/CA-substrates [1 µM], DMSO (1 % v/v) or remained untreated. After 1 h incubation at 37 °C, both dead cells (supernatant) and live cells (detached with trypsin) were collected. SYTOX Blue dead cell stain [1 µM] was added and the cells were subsequently analyzed by flow cytometry. Experiment was conducted in technical triplicates. a, Flow cytometry histograms of cells stained with SYTOX Blue, showing the gating strategy for live and dead cell events. b–d Dot plots showing the percentage of live cell events for b TMR-substrates, c CPY-substrates and d SiR-substrates in comparison to DMSO and untreated control samples. Each dot represents one replicate. The mean percentage of live cells from each triplicate is indicated as a black horizontal line. Error bars represent the s.d. The differences in cell viability of treated and untreated cells were not statistically significant (two-tailed unpaired t-test with Welch’s correction, p > 0.05, ns). Novel TF- and CF-substrates do not influence cell viability.
