Assessment of apoptosis in mESC as affect by 3BrP.

<p>ESCs were maintained in control conditions (with LIF), in the absence of LIF (differentiation control) and in the presence of two different 3BrP concentrations (25 and 50 μM) plus LIF. <b>A)</b>- Apoptosis/ Necrosis was accessed by flow cytometry for Annexin v/PI and three populations were identified live cells (negative for both annexin V and PI); early apoptotic cells (positive for annexin V and negative for PI); late apoptotic cells (positive for both annexin V and PI). At the left side of the panel representative dot plots for all experimental conditions are presented. 20000 cells were evaluated for Geometric Mean of Fluorescence in each population. 4 independent experiments were performed. Symbols are used to compare levels of live (*), early apoptotic (#) and late apoptotic (a) cells. One, two and three symbols represent p≤ 0.05, p≤ 0.01, p≤ 0.001, respectively. <b>B)</b>- Western Blot quantification by densiometric evaluation for p53 and caspase3. Results are presented as percentage relative to the CTR and were normalized for beta-actin. 3 independent experiments were performed. *p≤ 0.05, **p≤ 0.01, ***p≤ 0.001.