Figure 13

The effect of proteasome inhibitors on gene expression of TNF-α, and iNOS in LPS-stimulated peritoneal macrophages from LMP7/MECL-1^-/- knockout mice. Thioglycollate-elicited peritoneal macrophages were prepared from 8-week-old LMP7/MECL-1^-/- knockout mice, adhered to the bottom of 12 wells plate (1 × 10⁷ cells/well in 1 mL media) for 2 h, then treated with 100 μL dissolved in 0.2% DMSO of α-tocopherol (100 μM), δ-tocotrienol (10 μM), riboflavin (40 μM), or quercetin-HCL (40 μM) for 2 h. After 2 h, the cells were challenged with LPS + IFN-γ (10 ng/well; 400 μL + 50 units/mL), and incubated at room temperature for 4 h. Total cellular RNA was extracted and real-time polymerase chain reaction (RT-PCR) was conducted to quantitate expression of TNF-α (A), and iNOS (B) genes. Cell viability exceeded > 95%) in all the treatments. Data are presented as the percent of mRNA for each of the genes analyzed compared to controls. The treatments 1-5 correspond to: The treatments 1-5 correspond to: 1. Control (media + cells + LPS + 0.2% DMSO); 2. α,-tocopherol; 3. δ-tocotrienol; 4. riboflavin; 5. quercetin-HCL.