Gene expression profiles of transcription factors induced by CC-4047 in CD34⁺ cells. (A) CD34⁺ cells were cultured with SCF plus IL-3 plus IL-6 as indicated in Figure 2A in the presence of CC-4047 (100 μM) or 0.1% DMSO (control), followed by extraction of total RNA at day 1 (□), day 2 (), and day 3 (▪). Six micrograms of total RNA was subjected to DNA microarray analysis (see “Materials and methods”). The relative expression levels of transcription factors induced by CC-4047 are shown as percentage of increase or decrease (-) compared with control. (B) CD34⁺ cells were cultured with SCF plus IL-3 plus hyper–IL-6 (Figure 2A) in the presence of thalidomide (10 μM), CC-4047 (10 μM), or 0.1% DMSO (control), followed by extraction of total RNA (day 3) or protein lysates (day 7). RNA was analyzed for GATA-1 expression by Northern blotting. Methylene blue staining of 18S ribosomal RNA is shown as loading control. (C) The cell lysates were subjected to Western blotting to determine GATA-1 protein expression. β-actin expression served as loading control.