GFP-GATA2 fusion protein expression. (A) Schematic diagram of GFP-GATA2, GATA2-GFP, and GFP expression constructs. (B) Expression of GFP fusion proteins and DNA content of 293T cells at 24 hours after transfection. Black and gray histograms represent DNA contents of GFP⁺⁺ cells and total cells, respectively. Numbers indicate the percentage of cells in 6 rectangle regions. (C) 293T cells transfected with GFP-GATA2 fusion expression plasmids were stained with anti–cyclin B antibody (red). Cyclin B1 accumulates in the cytoplasm in late S phase, and later enters nuclei in M phase. GFP-GATA2 fusion proteins are expressed in cells in which the cytoplasmic stains for cyclin B1 and in cells expressing cyclin B1 with nuclear-envelope breakdown (white arrow). Thus, the fusion protein is expressed in late S phase and M phase. Original magnification × 630. (D) Cells transfected with GFP-GATA2–expressing plasmids (•) and with GFP-expressing plasmids (♦) were replated in separate dishes at 8 hours, and then the percentage of GFP-positive cells and GFP fluorescence intensity (Y geometric mean channel) were analyzed at 24, 48, and 72 hours after transfection.