Table 1.
| Primers | Sequence | Amplicon (pb) | |
|---|---|---|---|
| RS18 | forward | 5’-GCAGAATCCACGCCAGTACAA-3’ | 208 |
| reverse | 5’-GCCAGTGGTCTTGGTGTGCT-3’ | ||
| RPL32 | forward | 5’-CATTGGTTATGGAAGCAACAAA-3’ | 150 |
| reverse | 5’-TTCTTGGAGGAAACATTGTGAG-3’ | ||
| EBP | forward | 5’-TCCAGACATTACCTGGCAGCT-3’ | 111 |
| reverse | 5’-ATGTTGCTGCCTGCACTGTT-3’ | ||
| β-gal | forward | 5’-GCTGGTTATCCTGAGGCCC-3’ | 104 |
| reverse | 5’-CGGAGGAGCGGAGAAGAATA-3’ | ||
| NEU-1 | forward | 5’-AATGCCCGAAACCAGAACAAC-3’ | 239 |
| reverse | 5’-CGCCATGAGGTACCATTGCT-3’ | ||
| PPCA | forward | 5’-AATCTCTATGCCCCGTGTGCT-3’ | 102 |
| reverse | 5’-TGGCAGGCGAGTGAAGATG-3’ | ||
All primers were synthetized by Eurogentec (Angers, France). Real-time PCR was performed using an Absolute SYBR Green Rox mix (Thermo Electron, Courtaboeuf, France), and the Chromo Four-Color-Real-Time PCR detection system (Bio-Rad, France). PCR conditions were 15 minutes at 95°C, followed by 40 cycles each consisting of 15 s at 95°C (denaturation) and 1 minute at 60°C (annealing/extension). PCR efficiency of the primer sets (EBP, β-Gal, Neu-1, PPCA, RS18, and RPL32) was controlled via the slope of a standard curve. Results were standardized to RS18 and RPL32 gene expression levels using Genex software (BioRad).