Background
Methods
Cell culture
Animals and tissues collection
RNA extraction and RT-PCR
Gene names | Accession No. | Primers | Product | |
---|---|---|---|---|
GAPDH
| NM_001034034.1 | Forward | CCTTCATTGACCTTCACTACATGGTCTA | 173 - 200 |
Reverse | GCTGTAGCCAAATTCATTGTCGTACCA | 1029 - 1003 | ||
bCSH1
| NM_181007.2 | Forward | CCATCTCCCCATCAGCAGCAGT | 105 - 126 |
Reverse | GAGACCCATTACACCCAAACAT | 975 - 954 | ||
bPRP-1
| NM_174159.2 | Forward | CACGGTCAACAGGAGTCCTCACC | 43 - 65 |
Reverse | AATTTCAGGTAGCCCGCTGTGG | 873 - 852 | ||
bPAG1
| NM_174411.2 | Forward | CACCATTGGAACACCCCC | 245 - 262 |
Reverse | CACTGGGTAGTTGATGCCGTT | 989 - 969 | ||
IFNT
| NM_001015511.2 | Forward | TCCCCATGGCCTTCGTGCTCTCTCT | 75 - 99 |
Reverse | CTCAAAGTGAGTTCAGATCTCCACC | 668 - 644 | ||
bERVE-A
| NW_001493691.1 | Forward | TTCCTCAAAGAAGAAGAGGTAGAACAA | 703277-703304 |
Reverse | GGGTCCAAATAAGAGGAATAGAATGAT | 703748-703775 | ||
bERVE-B
| NW_001493674.2 | Forward | GAACTTAATGAGGATATGGAGCAGGTA | 3100208-3100181 |
Reverse | GACGTTTTGGGTAATCTTTAGTTGAGA | 3099456-3099429 | ||
BERV-K1 env | NW_001495384.1 | Forward | GGATGCAATTTGGATCCCAGAC | 218834-218855 |
Reverse | CCTTTGCATATTAGGCCTCTCCG | 221044-221022 | ||
BERV-K2 env | NW_001494209.3 | Forward | TCCACAGGACGCAGATTCTCC | 1926147-1926167 |
Reverse | CCTTTGCGTATGCCGAGCCTCCT | 1928389-1928367 |
Quantitative real-time RT-PCR (qRT-PCR)
Gene names | Accession No. | Primers | Product | |
---|---|---|---|---|
GAPDH
| NM_001034034.1 | Forward | AAGGCCATCACCATCTTCCA | 280 - 299 |
Reverse | CCACCACATACTCAGCACCAGCAT | 355 - 332 | ||
bCSH1
| NM_181007.2 | Forward | TGCCACACCGAATTCATGAC | 447 - 466 |
Reverse | AGGGCTTCGTCCTCTGTATTTG | 514 - 493 | ||
bPRP-1
| NM_174159.2 | Forward | CACGGAGCTGCAGCATATGA | 501 - 520 |
Reverse | CCTTGTGGCGCTTGATAGGA | 558 - 539 | ||
bPAG1
| NM_174411.2 | Forward | TCCACTTTCCGGCTTACCAA | 375 - 394 |
Reverse | CCTTTCATTCTCCCAGATCCAT | 436 - 415 | ||
IFNT
| NM_001015511.2 | Forward | CTAGGTGCCAGGCAGAACCT | 173 - 192 |
Reverse | GGGATGAGGAGAGAGTCTGTTCA | 232 - 210 | ||
bERVE-A
| NW_001493691.1 | Forward | GGATCTGACGGGAGACACAAA | 703321-703301 |
Reverse | CACCAATCCGGGAATCTTCA | 703281-703260 | ||
bERVE-B
| NW_001493674.2 | Forward | GGCCCAAGCACTCCTTCAT | 3100010-3099992 |
Reverse | CGCCCTTTTTCCCATTTCTT | 3099973-3099954 | ||
BERV-K1 env | AB587259.1 | Forward | GGAAATCACCGGGATGTCCT | 221-240 |
Reverse | GGAGAGGAGGCGCTTACCTG | 404-385 | ||
BERV-K2 env | AB587260.1 | Forward | AAAGGAGGTCAGGCCGCCTG | 221-240 |
Reverse | TGGGGGAGGAGGCGCTTACCT | 406-386 |
Induction of trophoblastic cell differentiation
Immunocytochemistry
Statistical analysis
Results
Expression profiles of ERV elements in the BT cell lines
Induction of binucleation and BNC-specific gene expression during on-matrigel culturing
Induction of the expression of ERV elements in BT cell lines grown by on-matrigel culturing
Correlation coefficients (r) | |||||||
---|---|---|---|---|---|---|---|
bPRP-1
|
bPAG1
|
IFNT
|
bERVE-A
|
bERVE-B
| BERV-K1env | BERV-K2env | |
bCSH1
| 0.94 ** | 0.92 ** | −0.48 | 0.94 ** | 0.26 | 0.92 ** | 0.59 |
bPRP-1
| 0.97 ** | −0.73 | 0.91 ** | 0.46 | 0.99 ** | 0.61 | |
bPAG1
| −0.70 | 0.94 ** | 0.34 | 0.94 ** | 0.57 | ||
IFNT
| −0.58 | −0.69 | −0.76* | −0.48 | |||
bERVE-A
| 0.22 | 0.89 ** | 0.62 | ||||
bERVE-B
| 0.50 | 0.71 | |||||
BERV-K1
env
| 0.59 | ||||||
BERV-K2
env
|