Table 3. Lineage-specific and overall changes in proliferation in response to RA and EDN3.
| Comparison of mean proportion of proliferating cells (p value) |
||||
|---|---|---|---|---|
| %EDU+/DAPI+ | %EDU+peripherin+/EDU+ | %EDU+SMA+/EDU+ | %EDU+S100β+/EDU+ | |
| RA- vs. RA+ (EDN3-) | 25.8 vs. 45.1 (p<0.001) | 8.8 vs. 8.4 | 28.5 vs. 18.3 | 35.9 vs. 63.8 (p<0.001) |
| RA- vs. RA+ (EDN3+) | 33.0 vs. 48.8 (p<0.001) | 3.5 vs. 3.5 | 19.2 vs. 8.4 | 14.4 vs. 45.4 (p<0.001) |
| EDN3- vs. EDN3+ (RA-) | 25.8 vs. 33.0 (p=0.014) | 8.8 vs. 3.5 | 28.5 vs. 19.2 | 35.9 vs. 14.4 (p=0.006) |
| EDN3- vs. EDN3+ (RA+) | 45.1 vs. 48.8 | 8.4 vs. 3.5 | 18.3 vs. 8.4 | 63.8 vs. 45.4 (p=0.015) |
| Interaction present? | No | No | No | No |
This table summarizes the effects of RA and EDN3 on the proliferation of ENS precursor cultures after 3 days. Immunoselected cells were cultured in the presence of RA, EDN3, RA with EDN3, or neither compound. When EDN3 was absent, the EDN signaling inhibitor BQ-788 was added to inhibit endogenous EDNRB signaling. The overall proportion of proliferating cells (EDU+/DAPI+), and the proportion of proliferating cells of each lineage as a percentage of all proliferating cells (EDU+) were quantified and compared between the groups. A two way ANOVA was employed to identify the contribution of each compound to changes in the proportion of proliferating cells. Significant changes (p<0.05) are in bold. Abbreviations: SMA =α-smooth muscle actin; DAPI = 4',6-diamidino-2-phenylindole; EDU = 5-ethynyl-2´-deoxyuridine; EDN3 = endothelin-3; RA = Retinoic acid;