Figure 1
Figure 1. GATA-1 represses VDR and is inversely expressed by myeloid DC subsets. (A) Schematic representation of culture models for LC and moDC differentiation used in this study. (B) Quantitative real-time RT-PCR analyses of VDR mRNA expression in GATA-1–transduced monocytic cells. U937Te cells were transduced with a retroviral vector encoding GATA-1-IRES-GFP or empty control vector (CTRL). GFP+ cells were sorted by FACS 48 hours after infection. mRNA was extracted from 105 cells. Cells are analyzed for VDR mRNA levels relative to HPRT mRNA (n = 3; *P < .05). (C) Representative Western blot analysis of VDR and GATA-1 protein levels in DC subsets (n = 3). CD34+-derived moDCs and LCs were generated from CD34+ cord blood cells; moDCs were generated from blood monocytes. (D) Fresh blood monocytes or monocytes during culture (days 2, 3, and 6) in the presence of GM-CSF/IL-4 (moDC cultures) were analyzed by Western blot (top) or by FACS (bottom; day 0, day 3, day 6). Each number in a quadrant represents the percentage of cells in the quadrant. Data are representative of 4 independent donors.

GATA-1 represses VDR and is inversely expressed by myeloid DC subsets. (A) Schematic representation of culture models for LC and moDC differentiation used in this study. (B) Quantitative real-time RT-PCR analyses of VDR mRNA expression in GATA-1–transduced monocytic cells. U937Te cells were transduced with a retroviral vector encoding GATA-1-IRES-GFP or empty control vector (CTRL). GFP+ cells were sorted by FACS 48 hours after infection. mRNA was extracted from 105 cells. Cells are analyzed for VDR mRNA levels relative to HPRT mRNA (n = 3; *P < .05). (C) Representative Western blot analysis of VDR and GATA-1 protein levels in DC subsets (n = 3). CD34+-derived moDCs and LCs were generated from CD34+ cord blood cells; moDCs were generated from blood monocytes. (D) Fresh blood monocytes or monocytes during culture (days 2, 3, and 6) in the presence of GM-CSF/IL-4 (moDC cultures) were analyzed by Western blot (top) or by FACS (bottom; day 0, day 3, day 6). Each number in a quadrant represents the percentage of cells in the quadrant. Data are representative of 4 independent donors.

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