Repolarization of protumoral M-MØ toward antitumoral MØ. (A) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses of M2 and M1 genes from M-MØ treated for 24 hours with GM-CSF (1000 U/mL), the MIF inhibitor 4-IPP (50 μM), or in combination. Values of M-MØ in the absence of treatment are given an arbitrary value of 1. Results represent mean ± SEM of 10 independent donors. (B) Flow cytometry histograms showing cell surface expression of FRβ, CD163, and ICAM-3 in M-MØ untreated or treated as indicated. Results from a representative MØ donor (upper graphs), and MFI ± SEM quantification of at least 4 independent experiments with the indicated treatments (lower graphs) are shown. Values in the absence of treatment are given an arbitrary value of 1. (C) Immunoblot analysis of phospho-AMPK (P-AMPK) expression in M-MØ untreated or treated for 6 hours with GM-CSF, 4-IPP, or in combination. Densitometric analyses (a.u.) normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels and referred to M-MØ control are shown. (D) Determination of NCI-H929 and U266 cell death alone (first bar) or cultured with M-MØ untreated or treated as indicated. 4-IPP (50 μM), M-CSF neutralizing Ab (1 μg/mL), M-CSFr inhibitor GW2580 (1 μM). Results represent mean ± SEM of 3 independent experiments with different donors. *P < .05; **P < .01; ***P < .001.