Figure 2
Increased membrane leakage followed in an IncuCyte live-imaging device. B76 cells were seeded in 96-well plates, IT and/or CsA were added and the fluorescence measured every second hour for 48 h (A). Membrane leakage (A) was quantified using the built in fluorescent object count metric after adding the dsDNA-binding YoYo-1 together with IT and/or CsA (n = 6 wells). Filled markers are used for CsA (diamond) and combinations of CsA and IT (triangle, CsA + IT 10 ng/ml and circle, CsA + IT 1 ng/ml). Open markers for control (diamond) and IT 100 ng/ml (square), 10 ng/ml (triangle), 1 ng/ml (circle). Results were filtered and fluorescent objects with an area > 100 μm2 is shown. The cytotoxic index (B) is the ratio of fluorescent objects before and after lysis of the cells by adding Triton X-100. The average with SD of 4 independent experiments each with six wells for each treatment is shown. The CI index obtained for untreated cells was subtracted in each experiment.