I am trying to characterize the ATM and p53 responses in a number of primary cell lines I have generated. I have been using western blot for pospho-p53 and p21 induction before and after irradiation.
However, I was looking at some ATCC recommended cell lines with mutant p53, and they have p21 induction after irradiation. What else can activate p21?
you can also try to determine acetylated p53 level. the acetylation status confirms p53 activation.
p21 can also be activated in p53 independent way. Di Fazio et al. Cellular Oncology 2010
Usually staining for p53 induction and p21 induction should be pretty reliable.
But then depends what kind of activation or activity you are looking for, as p53 can do different things in a cell, depending on the stimulus by which it got activated or induced. E.g. different phosphorylation sites, acetylation, level of expression...
However, as you and others mentioned above there are p53 independent ways of p21 induction. Another one is treatment with HDAC-inhibitors (like SAHA). These alter chromatin structure and thus allow p21 transcription.
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