Figure 11
The effect of proteasome inhibitors on gene expression of TNF-α, and iNOS in LPS-induced peritoneal macrophages from C57BL/6 mice. Thioglycollate-elicited peritoneal macrophages were prepared from 8-week-old C57BL/6 mice, adhered to the bottom of 12 well plates (1 × 107 cells/well in 1 mL media) for 2 h, after 2 h, 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. Then cells were challenged with LPS (10 ng/well; 400 μL), 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. Data are presented as the percent of mRNA for each of the genes analyzed compared to controls. The treatments 1-5 correspond to: 1. Control (media + macrophages (cells) + LPS + 0.2% DMSO); 2. α-tocopherol; 3. δ-tocotrienol; 4. riboflavin; 5. quercetin-HCL.