miRNAs-19b, -29b-2* and -339-5p Show an Early and Sustained Up-Regulation in Ischemic Models of Stroke (original) (raw)
Figure 2
OGD induced cell death demonstrates characteristics of apoptotic cell death.
A. The pan-caspase inhibitor, zVAD.fmk at a concentration of 100 µM, was able to significantly reduce levels of OGD-induced cell death from 70.6±2%, in naïve cells, to 54.3±5% in zVAD.fmk treated cells (p = 0.0459, n = 3) at 24 h post-OGD termination, thus indicating cell death is caspase dependent. B. Western blot analysis shows that OGD induces cytochrome C release into the cytosolic fraction of cell lysates. OGD also induces activation of Caspase 3, as indicated by the presence of the cleaved 17 kDa fragment. OGD-treated rat cortical neurons display PARP that has been cleaved to form the classic 89 kDa apoptosis fragment. The 50 kDa fragment of PARP that is found during necrosis is not detected. C. Quantification of the Western blots demonstrates an increase of cytochrome C intensity of 0.9±0.3, in controls, to 21±3.7 following OGD (p = 0.0056, n = 3). Quantification of Caspase 3 signal intensity shows a significant increase in OGD samples reaching 36.6±5.4 (p = 0.0046, n = 3). Quantification revealed that levels of the 89 kDa PARP fragment changed from 13.3±11 in control cells, to 135.6±16 in OGD treated rat cortical neurons (p = 0.0032, n = 3).Data represent Mean ± SEM.