Fig. 1.

NR4A3 functions as a tumor suppressor gene in human acute myeloid leukemia (AML) cells. (A) Quantitative real-time PCR analysis of NR4A3 expression in AML cell lines, human primary AML samples and CD34⁺ bone marrow mononuclear cells (BMMNC) from control subjects (Control-3, Control-4 and Control-5). mRNA levels were normalized to GAPDH expression. The expression level in CD34⁺BMMNC is shown as the mean ± S.D. (n = 3). The expression level in AML cell lines and human primary AML samples relative to the mean level in CD34⁺BMMNC is represented as the mean ± S.D. for triplicate analyses. (B-D) Kasumi-1, THP-1 and MOLM-13 cells were transduced with either the control or NR4A3 retrovirus and were purified for the experiments by cell sorting using green fluorescent protein (GFP) as a marker 72 h after infection. (B) NR4A3 expressions 72 h after GFP sorting were detected by Western blotting using the anti-FLAG antibody (upper panel) and the anti-NR4A3 antibody (lower panel). (C) The growth of the AML cells transduced with NR4A3. The data are shown as the means ± S.D. for triplicate analyses. (D) Colony formations in methylcellulose were determined for AML cells transduced with NR4A3. Total colonies were scored on day 10. The percentage of colony numbers relative to the controls is shown as the means ± S.D. for triplicate analyses. *, P < 0.05; **, P < 0.01