Clonal analysis of LacZ-positive cells after differentiation induction
| Promoter* . | Percentage of LacZ+ Cells . | ||||
|---|---|---|---|---|---|
| Megakaryocytes . | Erythrocytes . | Macrophages . | |||
| Day 8 . | Day 12 . | Day 8 . | Day 12 . | Day 10 . | |
| GPIIb (−598/+33) | 97.7 62.0 | 42.8 18.2 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 69.5 | 23.6 | <0.1 | <0.1 | <0.1 | |
| 85.8 | 37.4 | <0.1 | <0.1 | <0.1 | |
| 73.4 | 11.8 | <0.1 | <0.1 | <0.1 | |
| 78.0 | 63.5 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−400/+33) | 23.8 <0.1 | 53.4 0.5 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 5.8 | 9.6 | <0.1 | <0.1 | <0.1 | |
| <0.1 | 1.0 | <0.1 | <0.1 | <0.1 | |
| 0.5 | <0.1 | <0.1 | <0.1 | <0.1 | |
| 0.5 | <0.1 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−100/+33) | 0.9 4.7 | <0.1 6.3 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 2.0 | 0.9 | <0.1 | <0.1 | <0.1 | |
| 2.9 | <0.1 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−598/−400: −100/+33) | 48.5 72.2 58.3 | 31.1 18.2 14.1 | <0.1 <0.1 <0.1 | <0.1 <0.1 <0.1 | <0.1 <0.1 <0.1 |
| 27.2 | 3.7 | <0.1 | <0.1 | <0.1 | |
| 20.0 | 1.0 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−118/−93 m) | 98.6 3.8 | 96.5 1.9 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 33.5 | 8.2 | <0.1 | <0.1 | <0.1 | |
| 5.8 | 2.5 | <0.1 | <0.1 | <0.1 | |
| 11.4 | 2.1 | <0.1 | <0.1 | <0.1 | |
| 1.8 | 0.5 | <0.1 | <0.1 | <0.1 | |
| 26.8 | 3.0 | <0.1 | <0.1 | <0.1 | |
| 44.6 | 10.2 | <0.1 | <0.1 | <0.1 | |
| 6.5 | 0.9 | <0.1 | <0.1 | <0.1 | |
| EF-1α | ND | 3.5 | 3.4 | 0.2 | 7.0 |
| ND | 28.0 | 22.2 | 2.6 | 19.5 | |
| ND | 49.0 | 52.4 | 5.8 | 55.0 | |
| 6.3 | 9.7 | 5.5 | 1.1 | 2.9 | |
| 6.0 | 6.2 | 0.9 | 0.2 | 4.1 | |
| 25.1 | 32.1 | 10.3 | 6.5 | 20.4 | |
| 3.2 | 2.7 | 1.9 | 0.4 | 0.9 | |
| 32.5 | 32.4 | 33.1 | 9.3 | 20.2 | |
| Promoter* . | Percentage of LacZ+ Cells . | ||||
|---|---|---|---|---|---|
| Megakaryocytes . | Erythrocytes . | Macrophages . | |||
| Day 8 . | Day 12 . | Day 8 . | Day 12 . | Day 10 . | |
| GPIIb (−598/+33) | 97.7 62.0 | 42.8 18.2 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 69.5 | 23.6 | <0.1 | <0.1 | <0.1 | |
| 85.8 | 37.4 | <0.1 | <0.1 | <0.1 | |
| 73.4 | 11.8 | <0.1 | <0.1 | <0.1 | |
| 78.0 | 63.5 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−400/+33) | 23.8 <0.1 | 53.4 0.5 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 5.8 | 9.6 | <0.1 | <0.1 | <0.1 | |
| <0.1 | 1.0 | <0.1 | <0.1 | <0.1 | |
| 0.5 | <0.1 | <0.1 | <0.1 | <0.1 | |
| 0.5 | <0.1 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−100/+33) | 0.9 4.7 | <0.1 6.3 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 2.0 | 0.9 | <0.1 | <0.1 | <0.1 | |
| 2.9 | <0.1 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−598/−400: −100/+33) | 48.5 72.2 58.3 | 31.1 18.2 14.1 | <0.1 <0.1 <0.1 | <0.1 <0.1 <0.1 | <0.1 <0.1 <0.1 |
| 27.2 | 3.7 | <0.1 | <0.1 | <0.1 | |
| 20.0 | 1.0 | <0.1 | <0.1 | <0.1 | |
| GPIIb (−118/−93 m) | 98.6 3.8 | 96.5 1.9 | <0.1 <0.1 | <0.1 <0.1 | <0.1 <0.1 |
| 33.5 | 8.2 | <0.1 | <0.1 | <0.1 | |
| 5.8 | 2.5 | <0.1 | <0.1 | <0.1 | |
| 11.4 | 2.1 | <0.1 | <0.1 | <0.1 | |
| 1.8 | 0.5 | <0.1 | <0.1 | <0.1 | |
| 26.8 | 3.0 | <0.1 | <0.1 | <0.1 | |
| 44.6 | 10.2 | <0.1 | <0.1 | <0.1 | |
| 6.5 | 0.9 | <0.1 | <0.1 | <0.1 | |
| EF-1α | ND | 3.5 | 3.4 | 0.2 | 7.0 |
| ND | 28.0 | 22.2 | 2.6 | 19.5 | |
| ND | 49.0 | 52.4 | 5.8 | 55.0 | |
| 6.3 | 9.7 | 5.5 | 1.1 | 2.9 | |
| 6.0 | 6.2 | 0.9 | 0.2 | 4.1 | |
| 25.1 | 32.1 | 10.3 | 6.5 | 20.4 | |
| 3.2 | 2.7 | 1.9 | 0.4 | 0.9 | |
| 32.5 | 32.4 | 33.1 | 9.3 | 20.2 | |
Hematopoietic cells were generated from ES cells by the addition of TPO, EPO, or M-CSF for megakaryocytes, erythroid cells, and macrophages, respectively. ES cell clones that gave rise to LacZ-positive megakaryocytes were chosen randomly and the percentages of LacZ-positive cells relative to the total cells belonging to each lineage were calculated. Various mutated GPIIb promoters and the control EF-1α promoter were analyzed. The names of the GPIIb promoters are described in the text. ND indicates not determined.
The numbers in parentheses indicate the region of the GPIIb promoter. For example, −598/−400:−100/+33 indicates that a reporter construct consisting of nucleotides −598 to −400 and −100 to +33 of the GPIIb promoter was analyzed.