While studying a particular pathway under an induced metabolic state, I have noticed that the protein expression levels often increase in 6 hrs, decrease in 12 hrs and then subsides to normal level by 24 hrs and later. Is this how cells try to adapt to a particular metabolic condition? I have noticed this pattern in cyclins as well as mitochondrial dynamic proteins.
While going through previous researches on similar lines, I have seen that proteins of a particular pathway show a trend, either they increase with time or decrease, but for me, almost all of the proteins that i study show this increase, decrease and normal levles as compared to control.
Can anybody throw some light that can help me to draw a conclusion?
Are you comparing levels of these proteins to a housekeeping protein whose level is expected to remain constant? This is to control for variation in sample preparation.
Can you elaborate as what is your model system? And whether you are using any inducer for protein expression?
Adam B Shapiro I am comparing the levels with control. as in cells without my treatment but vehicle for the same time point. Housekeeping gene (beta actin) is my loading control.
Ankur Bothra I am using cell lines (osteoblasts). No I am not using any inducer. I am just treating the cells with my compound, and comparing the levels of proteins with expression with that of control at different time points.
Can you elucidate, what proteins are you working with? May they metabolize the compound? Respond to the application?
This depends on your experimental style, when using a singe dose setting your metabolic stimulus may lose its effects on your proteins of interest, this may led to this dynamic pattern, which is in general a normal reaction.
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