rhMIF induces VCAM-1 and ICAM-1 on MNs via Src and NFκB as determined by immunofluorescence. MNs were plated in 8-well chamber slides overnight. MNs were stimulated with rhMIF (50 nM) for 10 hours and studied for VCAM-1 and ICAM-1 expression using FITC-conjugated and PE-conjugated secondary antibodies. VCAM-1 and ICAM-1 expression was visible on the cell surface (A). To investigate Src activation by rhMIF, MNs were stimulated for 20 minutes. Panels B and C show immunopositivity for phospho-Src and phospho-NFκBp65 compared with NS cells. DAPI was added to stain nuclei. We have merged the DAPI and phosphorylation images in panels B and C to show the nuclear and cytoplasmic localization of phospho-NFκBp65 and phospho-Src. NS indicates nonstimulated. Images were captured using an Olympus BX fluorescence microscope with attached Olympus camera (Olympus, Melville, NY) and a 100 ×/1.3 numeric aperture objective, with oil as an imaging medium. Images were assembled using Adobe Photoshop software, version 7.01 (Adobe Systems, San Jose, CA).