Effect of Egr-1 on macrophage/granulocyte differentiation markers of myeloid-enriched BM cells.

Uninfected BM cells (BM control, □), mock-infected BM controls (BM mock, ), and BM infected with either MSCVneo (BMneo, ░) or MSCVneo Egr-1 (BMEgr-1, ▨) were seeded in methylcellulose supplemented with IL-3 (10% WEHI-3B–conditioned medium) with or without G418 (650 μg/mL). After 8 days, colonies were isolated, pooled, and washed, and the cells were resuspended in PBS. (A) Cells were used to prepare cytospin smears to assay for NSE staining and NBT reduction. Values are mean (± SD) results from 3 independent experiments. (B) For FACS analysis, cells were stained with FITC granulocyte-specific Gr-1 antibodies (antimouse Ly-6G) or macrophage-specific F4/80 antibodies (rat antimouse macrophage). Cell scattering and the intensity of fluorescence staining are shown. Three independent experiments were performed; results were similar.