Figure 2.

Activated PARP-1 leads to loss of MOMP mediated chromatinolysis. (a) AEBP1 silenced U138MG glioma cells (Lanes 6–9) show accumulation of large 50Kb fragments while doxycycline treated HeLa cells (Lane 3) show typical nucleosomal ladder pattern characteristic of apoptosis. (b) FACS analysis shows accumulation of sub n population of cells in AEBP1 down-regulated U138MG cells. AEBP1 siRNA treated cells were labeled with PI (50 µg/ml) and were gated for the Sub G₀ population as P1 in a Pulse Area Vs Pulse Width of the PE- 561 nm laser. (c) PARP-1 activation was monitored with PAR polymer production by western blot analysis at various time points upon AEBP1 down regulation. Un-transfected and scrambled siRNA (ScSi) treated U138MG cells were used as controls. (d) FACS analysis of Mitotracker measured at 577 nm in a Count Vs. Pulse-Area graph. Cells with live mitochondria metabolize the chloro-methyl moiety converting to a red fluorescent dye which can be read at 577 nm. The number in the graph is an average of three independent experiments indicating the percentage of cells showing signal at given time point and within the threshold intensity.