Hi, I would like to ask a question, the cell treated with drug for 48h, and the cell viability was changed using cck-8 kit, so I think the p-akt is decreased in western, however, the p-akt was increased in dose-dependent, and also, the total akt was increased in dose- dependent, but beta-actin was similar. How can I explain the result?
It is important to note the ratio of phosphorylated Akt/ total Akt regardless of the alteration of total Akt expression levels.
It is widely accepted that p-Akt undergoes up-regulation when cancer cells increase their viability.
It looks like your treatment up regulates Akt production. If the ratio of p-Act and Act is similar with increasing dose of tested substance there is no increase in Act signaling. As Act is involved in so many processes in the cell you should characterize the way of cell death (if it happened; apoptosis of necrosis) and do a cell cycle assay to see if your treatment induces cell cycle arrest.
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