Berberine reverses free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKβ

Figure 4 Effect of Ber on the IKKβ Ser¹⁸¹ Phosphorylation and IKKβ protein expression in 3T3-L1 Adipocytes. Fully differentiated cells were cultured for 12 h in serum-free DMEM with 0.2% BSA and then they were, respectively, cultured for 24 h in DMEM containing 0.5 mmol/L PA and 1% BSA (Mod); or cultured for 24 and 48 h in DMEM containing 0.5 mmol/L PA, 10 &mgr;mol/L Ber, 1% BSA, BH24, BH48; or in DMEM containing 0.5 mmol/L PA, 1 &mgr;mol/L Ber, 1% BSA for 24 h and 48 h (BL24, BL48); or cultured in DMEM containing 0.5 mmol/L PA, 5 mmol/L aspirin, 1% BSA for 24 h and 48 h (As24, As48); or in DMEM containing 1% BSA for 24 h (Nor). IKKβ phosphorylation and IKKβ protein expression were determined in the whole cell lysate by immunoblotting with the phosopho-spcific IKKβ (Ser¹⁸¹) antibody and IKKβ antibody respectively. Each experiment was repeated three times. ^bP < 0.01, as compared with Mod group.