Figure 7
IKKβ promotes proteasomal degradation of IκBα, which negatively regulates the expression of inflammatory genes. (A), a representative Western blot data set shows the effects of inhibitors on IκBα protein degradation induced by LCB. Differentiating 3T3-L1 adipocytes treated for 30 minutes with LCB (1:150 cells:LCB) and with or without 50 μM SC-514 (IKKβ inhibitor) or 20 μM MG132 (proteasome inhibitor) were collected for Western blot analysis using anti-IκBα and anti-β-actin antibodies. The IκBα:β-actin protein intensity ratio at each treatment was calculated and expressed as the relative ratio in respect to the value from the UT (untreated), as shown in the chart. (B–E), qPCR results show the effect of SC-514 inhibitor on inflammatory gene expression (IL-6, iNOS, MCP-1/CCL2, and COX-2) induced by LCB. Differentiating 3T3-L1 adipocytes were treated for 3 hours with LCB (1:150 cells:LCB) in the presence or the absence of 50 μM SC-514 inhibitor. (F–J) are like (A–E), respectively, except that 1% v/v LCB-RM was used to induce IκBα protein degradation.
