Figure 3. rMIF induces ICAM-1 expression in endothelial cells and monocytes.

(A) HMEC-1 (1×10⁶) and (B) HUVEC (1×10⁶) endothelial cells were stimulated with rMIF (0.4 µg/mL) for 24 h. ICAM-1 and nuclei were stained with FITC conjugated anti-ICAM-1 antibody and DAPI, respectively, and observed using fluorescence microscopy at 400× magnification. Negative controls were treated with 0.9% saline (Mock). Thrombin-treated HMEC-1 cells were used as a positive control. THP-1 monocytic cells (2×10⁶) (C and D) were stimulated with rMIF (0.4 µg/mL) for 24 h. (C) RNA was extracted and ICAM-1 expression was analyzed using a semi-quantitative RT-PCR with specific primers for ICAM-1. The gene expression of GADPH was used as the internal control. (D) The expression of ICAM-1 was detected by flow cytometry analysis and compared to the control (without rMIF treatment). Flow cytometry analysis showed increased ICAM-1 expression in THP-1 monocytes following rMIF treatment.