MIF-1 regulates differentiation in other cell lines and primary patient material.

A) Sorted CD138+ U266 cells and B) NCI-H929 cells were plated and treated with increasing concentrations of 4-IPP for 24 h. Cells were then stained with CD138 and PI, and CD138 content of viable (PI negative) cells is plotted. T = 0 is CD138 status at time of plating (N = 3) *pC) Steady state MIF-1 levels of sorted CD138+ and CD138- RPMI 8226 cells as measured by RT-PCR. D, E, F) Sorted CD138+ (solid) and CD138- (dashed) cells from RPMI8226 (D), U266-B1 (E), and NCI-H929 (F) lines were plated and media harvested at 1, 4, 12, and 24 h. post plating for use in the MIF ELISA assay (N = 3). *pG) MM cells derived from human bone marrow aspirates from patient H, stained for CD138 and CD38 at time of harvest and analyzed by flow cytometry. Data is also plotted in panel H. H) BM derived MM cells were placed in culture +/- 4-IPP for 24 h., then were stained with CD38 and CD138. The CD138 status of the CD38+ gate is shown.