LPS induction of Egr-1 expression in human monocytes and THP-1 cells.

(A) Nuclear extracts were prepared from adherent human monocytes with or without LPS (100 ng/mL) stimulation for 2 hours. Proteins were separated by SDS-PAGE and Egr-1 detected by Western blotting using anti–Egr-1 antibody. (B) THP-1 cells were stimulated with LPS (10 μg/mL) for various times (0-2 hours), and cytoplasmic and nuclear extracts were prepared. Egr-1 was detected by Western blotting. (C) THP-1 cells were stimulated with LPS for various times (0-24 hours). Egr-1 was detected in nuclear extracts by Western blotting. (D) EMSAs were performed using nuclear extracts and a radiolabeled oligonucleotide containing an Egr-1 site. Recombinant human Egr-1 (R) was used as a control.