Hi fellow researchers, has anyone ever worked with AICAR to induce activation of AMPK in L6 skeletal muscle cells? So far I tried to activate it by adding AICAR (either 0.1mM or 1mM) in DMEM with 1% pen strep to L6 myotubes. This was done after serum starvation for about 24hours. However, the AMPK was not activated when I compared with the untreated group. Has anyone ever activated it in these types of cells?
As I'm sure you are aware, activation of AMPK (i.e., increased phospho-(thr172)AMPKalpha ) by AICAR treatment of L6 myotubes has been reported several times. For example: PMCID: PMC2988518 or PMC3390622. I would recommend following one of these, especially as related to timing of treatment.
One point to consider is that serum starvation has been shown to increase pAMPK in myotubes. Is your question whether the effects of AICAR and serum starvation are additive?
Jeffrey J Brault Thank you for your response and recommendations. We'll optimize the time of treatment following these and others in literature through a time point study.
Interesting, I'll read more on how serum starvation increases pAMPK and whether the effects are additive. This wasn't my question but will definitely look into this too.
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