Figure 3. Evaluation of p53 expression and modification in HUVEC exposed to Bleo and SS.

HUVEC were treated for 4 h with Bleo (10 ug/ml) in static condition ST or exposed to SS. Control cells (open bars); Bleo treated cells (striped bars). 1. A representative western blotting analysis (WB) of total ATM expression (ATM) and phosphorylation (pATM) is shown on the left. The experiments were performed in static culture (ST), in the presence or absence of Bleo (B) and or laminar SS (SS). The graph on the right depicts the average level of ATM phosphorylation determined by densitometric analysis of three independent experiments. Data are expressed in arbitrary units. *** = p<0.001. 2. The picture on the left is a representative western blotting analysis of HIPK2 expression in cells in cells cultured in static condition (ST), treated with Bleo (B) and/or exposed to a laminar flow (SS). The presence of an unspecific band is indicated by a star on the left. Total actin content is shown as loading control. The graph on the right is the average densitometric analysis of three independent experiments. * = p<0.01. 3. The picture on the left shows the result of a representative western blotting analysis revealing p53 acetylation (p53^K382Ac) and phosphorylation on Serine 15 (p53^pSer15) or Serine 46 (p53^pSer46) in cells cultured in static culture (ST), treated with Bleo (B) and/or exposed to laminar flow (SS). The graph on the right shows the average densitometric values expressed in arbitrary units and obtained by three independent experiments. Data are expressed as percent (%) of control (C). * = p<0.05. ** = p<0.01. *** = p<0.001. 4. The picture (left panel) shows the result of a representative western blotting analysis of p21^{waf1,cip1,sdi1} expression in cells cultured in static condition (ST), treated with Bleo (B) and/or exposed to a laminar flow (SS). The graph on the right is the average densitometric analysis of three independent experiments. ** = p<0.01.