I checked the apoptotic effect of a natural molecule on cell line. In my western result I do get over expressed PARP Clvd and caspase 3 positive at 24 hours after treatment. When i checked level of Casapse 8 by fluorescence it shows signal significantly increased only after 36 hours. I did not checked BCL family proteins.
How can i co-related and make conclusion of my experiment.
There are multiple pathways leading to caspase 3 activation. If you wanted to identify the specific pathway, you would have to investigate the expression and activity of other enzymes involved, such as caspase 9 or the BCL family. If you simply want to demonstrate apoptosis, however, then caspase 3 cleavage and activity is probably sufficient.
So is that ok to say that " There is alternative pathway involved in apoptosis which do not require the activation of caspase 8 at that point of tim and seems caspase 3 activation is due to BCL family proteins activation. Which also involves decrease in mitochondrial potential and cytochrome C increase..........
Assuming that you can demonstrate each of those claims, then yes, that's okay. It depends on the specifics of your project. There are many projects where it is enough to simply demonstrate that apoptosis occurs by measuring levels of cleaved caspase 3 or caspase 3 activity. If it is necessary that you demonstrate the exact pathway leading to apoptosis, then you would need to study and provide data on those elements.
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