Figure 4. Effect of Pb in cellular aggregate formation.

mESC were incubated with the indicated Pb concentrations during the complete process of CA formation. (A) After CA disaggregation, cell numbers were recorded. (B) Active caspase-3 determinations. Cells were treated with 0 (control) or 0.1 µMPb (Pb) during the 8 days of CA formation process. CAs were disaggregated every two days and active caspase-3 was measured in the cultures (Control, yellow bars; Pb, pink bars). The supernatant was taken every two days and active caspase-3 was measured in the cells in suspension (Control, green bars; Pb, blue bars). Neurons obtained from mESC neural differentiation 3 days after CA disaggregation were also analyzed (3DIV, orange bar). Staurosporine (St, red bars) was used as a positive control of active caspase-3 induced apoptosis. The experiments were performed in three independent cultures. (*) p< 0.05.