To figure out optimal experimental conditions for palmitate-induced insulin resistance, HUVECs were starved serum for 6 h and then treated with 0.25, 0.5, and 0.75 mM palmitate for 12 h, or 0.5 mM palmitate for 6, 12, and 18 h, followed by stimulation with 100 nM insulin for 10 min. Here, INS is linked to Insulin resistance.