Cell cycle analysis of erythroblasts expressing different signaling proteins
Bicistronic retrovirus construct . | G1 phase, % . | S phase, % . | G2/M phase, % . |
|---|---|---|---|
| IRES-GFP | 81.8 ± 4.1 | 7.8 ± 2.2 | 10.4 ± 2.5 |
| H-ras. V12-IRES-GFP | 53.9 ± 0.6 | 40.0 ± 0.6 | 6.1 ± 0 |
| ca.MEK-IRES-GFP | 53.9 ± 6.0 | 39.8 ± 5.6 | 6.2 ± 1.8 |
| ca.Akt-IRES-GFP | 65.3 ± 7.3 | 20.1 ± 7.6 | 14.5 ± 1.0 |
| ca.Rlf-IRES-GFP | 70.2 ± 5.0 | 24.5 ± 6.4 | 5.3 ± 2.2 |
Bicistronic retrovirus construct . | G1 phase, % . | S phase, % . | G2/M phase, % . |
|---|---|---|---|
| IRES-GFP | 81.8 ± 4.1 | 7.8 ± 2.2 | 10.4 ± 2.5 |
| H-ras. V12-IRES-GFP | 53.9 ± 0.6 | 40.0 ± 0.6 | 6.1 ± 0 |
| ca.MEK-IRES-GFP | 53.9 ± 6.0 | 39.8 ± 5.6 | 6.2 ± 1.8 |
| ca.Akt-IRES-GFP | 65.3 ± 7.3 | 20.1 ± 7.6 | 14.5 ± 1.0 |
| ca.Rlf-IRES-GFP | 70.2 ± 5.0 | 24.5 ± 6.4 | 5.3 ± 2.2 |
Total fetal liver cells were freshly isolated from E13.5 to E15.5 embryos, and TER-119− erythroblasts were purified. Purified erythroblasts were infected with different bicistronic retrovirus constructs and cultured for 1 day in vitro. Infected cells were sorted by FACS and cultured in erythroid differentiation medium (EDM) for another day. Cultured cells were then incubated in hypotonic PI solution and analyzed by flow cytometry; the data were analyzed by ModFit software for cell cycle distribution. Results of infected cells are presented here as percentage of total cells analyzed.