TABLE I.
Primer Sequences (5′ to 3′) for Gene-Expression Analysis by Reverse Transcription Polymerase Chain Reaction, Annealing Temperatures, Number of Polymerase Chain Reaction Cycles Performed, and Expected Size of Each Polymerase Chain Reaction Product
| Gene* | Primer Sequence | Annealing Temperature | No. of Cycles Performed | Product Size (no. of base pairs) |
|---|---|---|---|---|
| GAPDH | ||||
| Forward | CAAGGCTGAGAACGGGAAGC | 58°C | 26 | 194 |
| Reverse | AGGGGGCAGAGATGATGACC | |||
| COL1A2 | ||||
| Forward | GGCTCCTGCTCCTCTTAGCG | 58°C | 26 | 132 |
| Reverse | CATGGTACCTGAGGCCGTTC | |||
| CBFA1/RUNX2 | ||||
| Forward | ACTGGGCCCTTTTTCAGA | 53°C | 26 | 317 |
| Reverse | GCGGAAGCATTCTGGAA | |||
| ALP | ||||
| Forward | TCAGAAGCTCAACACCAACG | 55°C | 26 | 199 |
| Reverse | GTCAGGGACCTGGGCATT | |||
| Osteocalcin | ||||
| Forward | GCCTTTGTGTCCAAGC | 52°C | 28 | 315 |
| Reverse | GGACCCCACATCCATAG | |||
| PPARγ2 | ||||
| Forward | TGAATGTGAAGCCCATTGAA | 53°C | 26 | 161 |
| Reverse | CTGCAGTAGCTGCACGTGTT | |||
| LPL | ||||
| Forward | GAGATTTCTCTGTATGGCACC | 52°C | 26 | 276 |
| Reverse | CTGCAAATGAGACACTTTCTC | |||
| FABP4 | ||||
| Forward | TGGGCCAGGAATTTGACGAAGT | 59°C | 24 | 215 |
| Reverse | TCAACGTCCCTTGGCTTATGCT | |||
| SOX9 | ||||
| Forward | ACATCTCCCCCAACGCCATC | 58°C | 34 | 173 |
| Reverse | TCGCTTCAGGTCAGCCTTGC | |||
| COL2A1 | ||||
| Forward | GGAAACTTTGCTGCCCAGATG | 58°C | 34 | 167 |
| Reverse | TCACCAGGTTCACCAGGATTGC | |||
| Aggrecan | ||||
| Forward | TGCGGGTCAACAGTGCCTATC | 58°C | 36 | 182 |
| Reverse | CACGATGCCTTTCACCACGAC |
*
GAPDH = Glyceraldehyde 3-phosphate dehydrogenase. ALP = Alkaline phosphatase. PPARγ2 = peroxisome proliferator-activated receptors-γ2. LPL = Lipoprotein lipase. FABP4 = fatty acid binding protein 4.