If I understand your question correctly, you detected basal levels of activating phosphorylation of AKT in your control sample (without MOI) in an experiment involving some cell-type. This observation is consistent with basal activation of AKT in cultured cells. More experimental details could help in addressing the question.

Thanks for replying me. Sorry, it was phosphorylated TAK, one of the activators of MAP kinase pathway.

Yes, you understood correctly. I got phosphorylated from in cells without any treatment (0 MOI)

Like Divaker Choubey said, it is hard to comment without knowing the details of the experiment. Seeing a phospho-protein signal in your control is not necessarily a bad thing. Did the phospho-signal go up or down with higher MOI? What knockout cells are you using?

Is this observation replicated? If you repeat your westerns from scratch (i.e. start with a new series of infections, lyse cells and do westerns), do you observe the same thing? If it is, then TAK is phosphorylated in your basal condition. Maybe it is normal in your experimental context. Maybe it is not and there is something in your media/experimental setup that is causing this abnormal effect. If you cannot replicate this observation, you don’t need to worry about it. It could have been a one-off.

Unless we know the context and have more details, it could be due to a million reasons. Maybe you could look up TAK/AKT signaling pathway and see if there is something that explains this observation.

I agree with above comments. Have you starved your cells before harvesting them? Usually, it works well to decrease basal Akt activation,

Best,

No problems with your result)

Thanks for all responses. I will repeat the experiment and see what will happen again.