qPCR validation of microarray results
| Gene . | Fold change . | |
|---|---|---|
| qPCR . | Microarray . | |
| Smad1 morphants | ||
| Cyclin G1 | 2.97 | 2.21 |
| Dazl | 3.59 | 9.29 |
| Rap2 inter. protein | 39.3 | 28.6 |
| Runx3 | 0.159 | −2.23 |
| Kruppel-like factor 12 | 0.489 | −2.00 |
| Smaf2 | 0.349 | −2.55 |
| Wnt16 | 0.415 | −2.67 |
| Smad5 morphants | ||
| GDNF receptor α-1a | 1.51 | 2.65 |
| HoxC1a | 1.56 | 2.08 |
| Angiopoietin-like 3 | 3.38 | 3.58 |
| Dlx5 | 0.216 | −10.0 |
| Bmp4 | 0.300 | −5.68 |
| Gata2 | 0.482 | −3.13 |
| Gene . | Fold change . | |
|---|---|---|
| qPCR . | Microarray . | |
| Smad1 morphants | ||
| Cyclin G1 | 2.97 | 2.21 |
| Dazl | 3.59 | 9.29 |
| Rap2 inter. protein | 39.3 | 28.6 |
| Runx3 | 0.159 | −2.23 |
| Kruppel-like factor 12 | 0.489 | −2.00 |
| Smaf2 | 0.349 | −2.55 |
| Wnt16 | 0.415 | −2.67 |
| Smad5 morphants | ||
| GDNF receptor α-1a | 1.51 | 2.65 |
| HoxC1a | 1.56 | 2.08 |
| Angiopoietin-like 3 | 3.38 | 3.58 |
| Dlx5 | 0.216 | −10.0 |
| Bmp4 | 0.300 | −5.68 |
| Gata2 | 0.482 | −3.13 |
A total of 13 genes shown by the microarray experiments to have altered expression in morphant embryos were chosen for validation by quantitative real-time PCR. The qPCR reactions were carried out on cDNA from smad1 or smad5 morphant embryos at the 1-somite stage. Shown is the average fold change in expression calculated from 3 independent experiments, with samples analyzed each time in triplicate. Samples were normalized to β-actin, and wild-type was set to 1. In every case, the differences compared with wild-type are significant by Student t test; P < .05. Also shown is the fold change as determined by microarray with negative fold change indicating underexpression in the morphant samples.