Table 1. ISH probes: Primers, amplicon sizes and temperatures used in the hybridization assays.
Gene-specific forward (F) and reverse (R) primers were used to generate antisense RNA probes for use in the in situ hybridization (ISH) assays. All R primers also contained the T7 RNA promoter sequence (T7PS) at the 5’-end (5´-TAATACGACTCACTATAGGGAGA-3´). The amplicon sizes and temperatures used in the ISH assays are also shown.
| Gene | Primers | Amplicon size (bp) | Temperatures | Reference sequence (NCBI) | |
|---|---|---|---|---|---|
| Col2a1 | F | 5’-TGGTGACAAGGGAGAAAAGG-3’ | 764 | 56°C | NM_001113515.2 |
| R | 5’- T7PS AACCTTGAGCACCTTCAGGA-3’ | ||||
| Dcx | F | 5’-GGGGATTGTGTACGCTGTTT-3’ | 779 | 57°C | AF045547.1 |
| R | 5’- T7PS TTGAGAGCTGACTGCTGGAA-3’ | ||||
| Runx2 | F | 5’-CACTGCCACCTCTGACTTCT-3’ | 752 | 68°C | NM_001146038.2 |
| R | 5’- T7PS CCTTGGTAAAGGGGACATCT-3’ | ||||
| Sox9 | F | 5’-AGAGCGAGGAAGATAAGTTC-3’ | 454 | 58°C | NM_011448.4 |
| R | 5’- T7PS ATTAGGAGAGATGTGAGTCT-3’ | ||||