Fig. 5

Vps18^+/- mutant Hoxb8 neutrophil progenitors undergo premature apoptosis.

A, B Cells positive for active caspase-3 at day 3 of differentiation in % of all single cells in indicated genotypes analyzed by flow cytometry. n = 6. **P < 0.01, ****P < 0.0001 compared to CTRL (A) or compared to clone 1 (B). C Representative Western blot of caspase-3 expression (left panel) and quantitative analysis (right panel) of active caspase-3 expression in cell lysates from indicated cell lines at day 3 of differentiation. Ratios of active caspase-3/GAPDH are presented as relative protein amount. n = 5. Mean ± SEM. One-way ANOVA, Šídáks multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001 compared to CTRL. ns, not significantly different. D, E Representative dot plots (D) and quantification (E) of viable, early and late apoptotic cells of indicated genotypes during differentiation (day 0-4). Viable cells, TMRM⁺, blue. Early apoptotic cells, Annexin V⁺ and SytoxRed^-, red. Late apoptotic cells, Annexin V⁺ and SytoxRed⁺, black. Numbers indicate % of cells of all single cells (100%, sum of all single cells). n = 4. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared to d0. (A, B, E) Mean ± SEM, one-way ANOVA, Tukey’s multiple comparisons test.