hi everyone,
I have ben treating cells with a drug for 4 months now and then immunoblotted with AKT as it has been suggested that that drug effects the PI3k pathway. however, the phosphorylation of the treated cells was decreed compared to the control yet the cells were dividing and growing in the 4 months duration!
any ides on why is this sudden result!
thank you
There are many possibilities to explain what happened:
1) Dose used may have been insufficient.
2) Akt decreased phosphorylation may have not achieved real block of the pathway.
Suggestion
Use another Akt phosphorylation blocker as control instead of using non treated cells.
Have you also looked at the different AKT isoforms or did you use panAKT antibodies? In many papers, people don't pay attention to it and sometimes only one isoform is relevant.
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