Figure 5. rMIF induces thrombomodulin (TM) expression in monocytes.

(A) THP-1 cells (2×10⁶) were stimulated with rMIF (0.4 µg/mL) for 24 h. TM and nuclei were stained with FITC conjugated anti-TM antibody and DAPI, respectively, and observed using fluorescence microscopy at 400× magnification. The controls were treated with 0.9% saline (Mock). (B) Flow cytometry analysis of TM expression in THP-1 cells with/without rMIF treatment. A neutralizing anti-MIF mAb (20 µg/mL) was used to abrogate the MIF-induced TM expression. (C) RNA was extracted and TM expression was analyzed through a semi-quantitative RT-PCR with specific primers for TM. The GADPH was used as the internal control. TM expression was enhanced with rMIF treatment in THP-1 cells, compared to the control (c; without rMIF treatment). (D) Flow cytometry analysis also showed increased TM expression in PBMCs following rMIF treatment.