Table 1.
Blockade of RAR-α or of PKA inhibits the induction of Prox1+ cell clusters by RA and cAMP
| Day 16 |
Day 18 |
|||
|---|---|---|---|---|
| EBs1 | clusters2 | EBs1 | clusters2 | |
| Control | 0/9 | 0 | 0/9 | 0 |
| 10 μM RA | 0/9 | 0 | 0/9 | 0 |
| 10 μM RA + 0.5 mM cAMP | 9/9 | 3+1 | 9/9 | 1+0 |
| 10 μM RA + 0.5 mM cAMP + 10 μM Ro 41-5253 | 0/9 | 0 | 0/9 | 0 |
| 10 μM RA + 0.5 mM cAMP + 10 μM H89 | 0/9 | 0 | 0/9 | 0 |
| 0.5 mM cAMP | 0/9 | 0 | 0/9 | 0 |
| 10 μM Ro 41-5253 | 0/9 | 0 | 0/9 | 0 |
| 10 μM H89 | 0/9 | 0 | 0/9 | 0 |
Four days of treatment with RA and cAMP induced the formation of 3+1 Prox1+/LYVE-1+/CD31+ cell clusters per EB. These clusters were found in 9 of 9 EBs analyzed. Ro 41-5253 (RAR-α inhibitor) and H89 (PKA inhibitor) completely prevented the induction of Prox1 endothelial cell clusters by RA and cAMP. No induction was seen after incubation with RA alone, cAMP alone or with the inhibitors alone.
1
Number of analyzed EBs.
2
Average number of CD31+/LYVE-1+/Prox1+ cell clusters per EB + SD.