Fig. 2: Pharmacological inhibition of caspase-3 blocked apoptosis but not preceding compaction, whereas blockade of myosin activity induced non-apoptosis, necrosis-like cell death.
From: Chromatin compaction precedes apoptosis in developing neurons
Live cell confocal images were acquired every 10 min for 3 time points before the application of the pharmacological treatments (baseline), and for an additional 39 time points (i.e., 6.5 h) after application of indicated pharmacological treatments. a–c Normalized nuclear size, edge count and CCP for all neurons under control (black, n = 460 cells), staurosporine (1.5 µM; orange, n = 518 cells), BDM (20 mM) with staurosporine (blue, n = 301 cells), and caspase-3 inhibitor Z-DEVD-FMK (100 µM) with staurosporine (green, n = 265 cells) conditions. Note, that while the combined application of caspase-3 inhibitor with staurosporine blocked average caspase activation and reduced the nuclear size change, no significant differences in time course and extent of early chromatin compaction were found between caspase-3 inhibitor with staurosporine and staurosporine-only treated neurons. d Mean percentage of NucView-positive cells (caspase activation) per culture (control n = 9 cultures, staurosporine n = 9 cultures, caspase-3 inhibitor Z-DEVD-FMK with staurosporine n = 5 cultures, BDM with staurosporine n = 5 cultures) identified throughout the acquisition. e Caspase activity measured by luminescent caspase assay at DIV8 under baseline conditions, as well as 3 h and 6 h after start of pharmacological treatments. Data are represented as mean ± SEM. For a–d two-way ANOVA was applied to detect differences across pharmacological treatments during baseline: nuclear size F(3, 4620) = 0, p > 0.9999; edge count F(3, 4620) = 3.45e−13, p > 0.9999; CCP F(3, 4620) = 0.0, p > 0.9999; mean percentage of NucView-positive cells F(3, 24) = 0.000, and during the time period after application of pharmacological treatments: nuclear size F(3, 60060) = 906.2, p < 0.0001; edge count F(3, 60060) = 1526, p < 0.0001; CCP F(3, 60060) = 2344, p < 0.0001; mean percentage of NucView-positive cells F(3, 24) = 4.82, p = 0.0092. For e a Kruskal-Wallis test was applied: H(7) = 39.56, p < 0.001.
