SBI-477

SBI-477, that coordinately inhibits triacylglyceride (TAG) synthesis and enhances basal glucose uptake in human skeletal myocytes, stimulates insulin signaling by deactivating the transcription factor MondoA, leading to reduced expression of the insulin pathway suppressors thioredoxin-interacting protein (TXNIP) and arrestin domain containing 4 (ARRDC4).^[1]

Primary human skeletal myotubes were grown and differentiated in 24-well plates. Cells were   treated with the indicated concentration of SBI-477 for 24 h. Following compound treatment, cells were rinsed three times with PBS and then incubated in 125μM [³H]-palmitic acid (60 Ci/mmol) bound to fatty acid free albumin containing 1mM carnitine for 2 hours at 37℃. The cell medium was transferred to a tube containing cold 10% trichloroacetic acid (TCA). The tubes were centrifuged at 8,500 x g for 10 minutes at 4℃. The supernatant was immediately removed, mixed with 6N NaOH, and applied to ion-exchange resin. The eluate was collected, measured by liquid scintillation analyzer and normalized to total protein amount. The amount of cell protein was measured.