Figure 4.

GA treatment does not affect protein or mRNA levels of CD14. Cells (6 × 10⁶) were incubated with or without GA (1 μg/ml) for 16 h and incubated with LPS (100 ng/ml) for 3–5 h. Then, cells were lysed for protein analysis or for RNA extraction. (A) Western blot for CD14. Cell extracts (100 μg of protein) were separated by SDS-PAGE, transferred onto a nitrocellulose membrane and immunodetected using HRP-conjugated anti-CD14 antibody. (B) Northern blot for CD14 mRNA. Total RNA (10 μg) was separated in a formaldehyde-agarose gel, transferred onto nylon modified membrane and hybridized with a radiolabeled cDNA probe for CD14. These blots are representative of three independent experiments.