TABLE 1
The 6 most downregulated genes involved in regulation of granulosa cells differentiation towards endodermal and epithelial tissues were validated using RT-qPCR, 168h/0h
| GENE NAME | GENE SYMBOL | FOLD CHANGE 48H/0H | ADJ. P. VAL. |
|---|---|---|---|
| integral membrane protein 2A | ITM2A | -8.85 | <0.05 |
| disabled homolog 1 (Drosophila) | DAB1 | -8.91 | <0.05 |
| mal, T-cell differentiation protein 2 | MAL2 | -21.75 | <0.05 |
| nebulette | NEBL | -26.72 | <0.05 |
| death associated protein-like 1 | DAPL1 | -28.69 | <0.05 |
| hydroxysteroid (17-beta) dehydrogenase 1 | HSD17B1 | -35.75 | <0.05 |
TABLE 2
Primer sequences (5′-3′)
| GENE NAME | GENE SYMBOL | PRIMER SEQUENCES (5′-3′) |
|---|---|---|
| integral membrane protein 2A | ITM2A | TCTCGTAGGCCTTTCCTTCA AGGCAGGAAGTAGGGCTCTC |
| disabled homolog 1 (Drosophila) | DAB1 | TACGTTTGTGGGAAGGAAGG CTTCCTTCTTTTGGCTGGTG |
| mal, T-cell differentiation protein 2 | MAL2 | AGGATGGGTCATGTTCGTGT TTGTCATTCAAGAGCGGCTG |
| nebulette | NEBL | CAAACCCTTCAAGGCTACCA CTGAGAACACGCTTCCATCA |
| death associated protein-like 1 | DAPL1 | CCTGCTCTGGAGAAGGTCAC GGGCCTAAGGAAAGTTTTGG |
| hydroxysteroid (17-beta) dehydrogenase 1 | HSD17B1 | GTGTCAGAGGCTTGCTAGGG CAGCACAATCTCAAGGCTGA |
FIGURE 1
Heatmaps presenting differentially expressed genes involved in “developmental process”, “cellular developmental process”, “regulation of cell differentiation”, “epithelium migration”, “epithelial cell proliferation”, “epithelial cell migration”, “epithelial cell differentiation”, “epithelial cell development”, “endodermal cell differentiation”, “endoderm formation”, “endoderm development” and “epithelium development” based on GO BP terms. Each row on the Y axis represents a single transcript. The red color indicates downregulated genes while the green are upregulated
FIGURE 2
The circular scatter plots of differentially expressed genes involved in “developmental process”, “cellular developmental process”, “regulation of cell differentiation”, “epithelium migration”, “epithelial cell proliferation”, “epithelial cell migration”, “epithelial cell differentiation”, “epithelial cell development”, “endodermal cell differentiation”, “endoderm formation”, “endoderm development” and “epithelium development” GO BP terms. Each dot represents a single gene. The z-scores were presented as segments of inner circles
FIGURE 3
The dendrogram of differentially expressed genes involved in “developmental process”, “cellular developmental process”, “regulation of cell differentiation”, “epithelium migration”, “epithelial cell proliferation”, “epithelial cell migration”, “epithelial cell differentiation”, “epithelial cell development”, “endodermal cell differentiation”, “endoderm formation”, “endoderm development” and “epithelium development” GO BP terms. The DEGs were clustered based on their logFC values
FIGURE 4
Analysis of enriched gene ontological groups involved in regulation of granulosa cells differentiation towards endodermal and epithelial tissues. The network plot presenting the linkages of genes and GO BP terms
FIGURE 5
Heatmap presenting the relationship between genes and selected GO BP terms. The yellow color of tiles indicates the absence of logFC values
FIGURE 6
Reactome FI network for “positive regulation of cell proliferation”. “--->” indicates activating/catalyzing, “-|” for inhibition, “-” FIs extracted from complexes or inputs and “---” predicted FIs
FIGURE 7
Microarray validation using RT-qPCR