Effect of STAT5 isoforms on IL-3–cultured CD45+ cell proliferation
| Cell-cycle phases . | Circulating human CD45+ cells . | Mouse BM-derived CD45+ cells . | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Scramble . | siRNA STAT5A . | siRNA STAT5B . | SiRNA STAT5A+ STAT5B . | SU6656 . | Scramble . | siRNA STAT5A . | siRNA STAT5B . | siRNA STAT5A+ STAT5B . | |
| G0/G1 | 40 ± 2 | 55 ± 3 | 57 ± 1 | 64 ± 1 | 63 ± 2 | 52 ± 2 | 70 ± 3 | 69 ± 1 | 75 ± 2 |
| S | 50 ± 3 | 31 ± 3* | 33 ± 1* | 26 ± 2* | 28 ± 1* | 35 ± 2 | 17 ± 2* | 15 ± 2* | 13 ± 3* |
| G2/M | 10 ± 1 | 14 ± 1 | 10 ± 2 | 10 ± 3 | 9 ± 3 | 13 ± 3 | 13 ± 1 | 16 ± 3 | 12 ± 2 |
| Cell-cycle phases . | Circulating human CD45+ cells . | Mouse BM-derived CD45+ cells . | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Scramble . | siRNA STAT5A . | siRNA STAT5B . | SiRNA STAT5A+ STAT5B . | SU6656 . | Scramble . | siRNA STAT5A . | siRNA STAT5B . | siRNA STAT5A+ STAT5B . | |
| G0/G1 | 40 ± 2 | 55 ± 3 | 57 ± 1 | 64 ± 1 | 63 ± 2 | 52 ± 2 | 70 ± 3 | 69 ± 1 | 75 ± 2 |
| S | 50 ± 3 | 31 ± 3* | 33 ± 1* | 26 ± 2* | 28 ± 1* | 35 ± 2 | 17 ± 2* | 15 ± 2* | 13 ± 3* |
| G2/M | 10 ± 1 | 14 ± 1 | 10 ± 2 | 10 ± 3 | 9 ± 3 | 13 ± 3 | 13 ± 1 | 16 ± 3 | 12 ± 2 |
Circulating human CD45+ cells and mouse BM-derived CD45+ cells cultured in the presence of IL-3 were transiently transfected with scramble sequence or STAT5A and STAT5B siRNA, alone or in combination. After 48 hours of treatment, the cells were fixed with 70% ethanol. DNA was stained with propidium iodide and analyzed with a flow cytometer (FACScan; BD Immunocytometry Systems, San Jose, CA). The percentage of cells in each phase of the cell cycle was determined by ModFit LT software (Verity Software House, Topsham, ME). Human CD45+ cells were also pretreated with SU6656. The numbers are the means plus or minus SD of 3 different experiments done on separate days, each performed in duplicate. (*P < .05, experimental groups vs scramble.)