Page 70 - Haematologica May 2022

G. Purgatorio et al.
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haematologica | 2022; 107(5)
Figure 4 Impaired angiogenesis in blood outgrowth endothelial cells (BOEC) from GATA2-mutant patients and in GATA2-silenced control blood outgrowth endothelial cells is restored by exogenous supplementation with nitric oxide. Impaired capillary tube formation on matrigel-coated wells by blood outgrowth endothelial cells (BOEC) from healthy controls, the unaffected family member (I1) and the GATA2-deficient family members and GATA2-silenced BOEC in com- parison with healthy control BOEC and BOEC transfected with a scramble small interfering RNA (siRNA) (NC), respectively. Treatment with the nitric oxide (NO) donor SNAP (100 μM for 24 hours restored tube formation in BOEC from GATA2- mutated patients and in GATA2-silenced BOEC. Angiogenesis was quantified with ImageJ software (AngioTool 64) by measuring total tube length, tube number and branching points. Values are means ± standard error of the mean (SEM) of 6 repeated measures (*P<0.0001 vs. control and #P<0.0001 vs. vehicle, two-way ANOVA followed by Tukey’s multiple comparison test). Specimens were analyzed at room temperature by a Carl Zeiss Axio Observer.A1 microscope (Carl Zeiss Inc, Oberkochen, Germany) using a 2.5X Plan-Apochromat objective and images acquired using the AxioVision software (Carl Zeiss Inc).