Fig. 4

Serum starvation induces apoptosis in male ECs. a Representative flow cytometry graphs illustrating the percentage of early apoptotic cells (upper left quadrant (UL)), late apoptotic cells (upper right quadrant (UR)), and live cells (lower left quadrant (LL)) in serum-starved M-ECs and F-ECs (left and right panels, respectively). b Means ± s.e.m. of the percentage of gated M-ECs and F-ECs (blue and deep red bars, respectively) in the UL, UR, and LL quadrants from 4 to 5 independent experiments. *p < 0.05, **p < 0.01 vs F-ECs, n = 4–5. c Pro- and cleaved caspase-3 were evaluated by immunoblotting in M- and F-EC lysates (30 μg/lane) prepared after an overnight incubation in 2% FBS. Upper panel: a representative blot out of six. Lower panel: a densitometric analysis of the ratio between cleaved and pro-caspase-3 forms is shown. *p < 0.05, n = 6. d Cleaved PARP-1 was evaluated as in c. β-Actin was used as a loading control. Upper panel: a representative blot out of three. Lower panel: the ratio between cleaved PARP-1 and β-actin is shown. *p < 0.05, n = 3