We modelled the effect of hyperinsulinemia, free fatty acid and hyperglycemia on insulin signalling in cell culture by prolonged (16 h) treatment of C2C12 mouse myotubes with insulin (10 nM), saturated fatty acid (palmitate, 500 μM) or high glucose (25 mM, instead of 5.5 mM). Here, INS is linked to hyperinsulinism.