Table 2.
Sequences of quantitative RT-PCR primers utilized for analysis of mRNA expression
| Gene | Primers for qRT-PCR | qRT-PCR Characteristics | ||||
|---|---|---|---|---|---|---|
| PPIA | F | GGTTTATGTGTCAGGGTGGTG | R | TCCCCATAGATGGACTTGC | E = 2.00 ± 0.06 | Ct = 24.8 ± 1.5 |
| β2-m | F | GAGGCTATCCAGCGTACTCC | R | TCCATTCTCTGCTGGATGAC | E = 2.00 ± 0.04 | Ct = 22.6 ± 1.5 |
| RPL32 | F | TTAAGCGTAACTGGCGGAAAC | R | GGCCCTTGAATCTTCTACGAAC | E = 1.98 ± 0.13 | Ct = 21.1 ± 1.6 |
| PolR2A | F | GGAGATCTTCACGGTGCTG | R | AGCCATCAAAGGAGATGACG | E = 1.97 ± 0.05 | Ct = 29.2 ± 1.3 |
| B-actin | F | ACCCCGTGCTGCTGAC | R | AACATGATCTGGGTCATCTTCTC | E = 1.96 ± 0.05 | Ct = 25.5 ± 1.5 |
| MyHC1 | F | AGGAGCTCACCTACCAGACG | R | TGCAGCTTGTCTACCAGGTC | E = 1.93 ± 0.03 | Ct = 21.3 ± 1.5 |
| MyHC2A | F | AACATGAGAGGCGAGTGAAG | R | GTGTTGGATTGTTCCTCAGC | E = 1.83 ± 0.04 | Ct = 23.5 ± 1.5 |
| MyHC2X | F | TGGTGGACAAACTGCAAGC | R | TTGTTCCTCCGCTTCTTCAG | E = 1.79 ± 0.05 | Ct = 24.9 ± 1.7 |
| PGC1α s4 | F | TGTGCCATATCTTCCAGTGACC | R | TGCAGTTCCAGAGAGTTCCAC | E = 1.97 ± 0.07 | Ct = 24.8 ± 2.7 |
| Ankrd2 | F | AGAAGCTGCCCATGGACTTG | R | TTGGCCCTTCACCTTCTGC | E = 1.91 ± 0.04 | Ct = 25.5 ± 2.1 |
| FGF6 | F | TGGTGAGTCTCTTTGGAGTGAG | R | TTGTTGGGCAGGAGGGTTTC | E = 1.99 ± 0.05 | Ct = 27.4 ± 1.5 |
| IGF1Ea | F | ATGCCCAAGACCCAGAAGG | R | CATCCTGTAGTTCTTGTTTCCTG | E = 1.98 ± 0.10 | Ct = 28.7 ± 2.1 |
| β-catenin | F | TGGCAACCAAGAAAGCAAGC | R | ACAGATAGCACCTTCAGCACTC | E = 1.93 ± 0.09 | Ct = 26.3 ± 2.6 |
| Myostatin | F | CCATGCCTACAGAGTCTGATTTTC | R | AGAAGCAACATTTGGGTTTTCC | E = 1.95 ± 0.09 | Ct = 30.1 ± 2.9 |
| Follistatin | F | GTGCACTCCTAAAGGCAAGATG | R | CCGACAAGTCTTTTTACATCTGC | E = 1.83 ± 0.06 | Ct = 34.5 ± 1.7 |
| PAX7 | F | AGCTGGAGAAGGCCTTTGAG | R | TACTGAACCAGACCTGCACAC | E = 1.97 ± 0.05 | Ct = 31.5 ± 1.5 |
| MCAD | F | AGCGTCATCTACAGCATCCAG | R | AAACGCTCTTAGCCTGAAGC | E = 1.97 ± 0.05 | Ct = 30.1 ± 1.4 |
| ITGA7 | F | GGCTTTCCAGATATTGCAGTGG | R | TGCTCCCATGGTAGATGAAGAC | E = 2.00 ± 0.05 | Ct = 27.0 ± 1.4 |
| Jagged-1 | F | ACATGTGGCCATTTCTGCTG | R | CAATCAGCGAGCTGTTTCCATC | E = 1.97 ± 0.05 | Ct = 30.2 ± 1.2 |
| CAV-1 | F | CCTAAACACCTCAACGATGACG | R | TCGTCACAGTGAAGGTGGTG | E = 1.97 ± 0.04 | Ct = 26.0 ± 1.4 |
| MYOG | F | AGCGAATGCAGCTCTCACAG | R | AGATGATCCCCTGGGTTGG | E = 1.97 ± 0.05 | Ct = 28.4 ± 1.2 |
| MYOD | F | ACGGCATGATGGACTACAGC | R | TTGTAGTAGGCGCCTTCGTAG | E = 1.90 ± 0.05 | Ct = 31.8 ± 1.5 |
| MRF4 | F | ATAACGGCTAAGGAAGGAGGAG | R | AGAAAGGCATCGAAGGCTACTC | E = 1.99 ± 0.04 | Ct = 27.1 ± 0.9 |
| MYF5 | F | AACTGCTCTGATGGCATGC | R | AGTACTGCTCTTTCTGGACCAG | E = 1.96 ± 0.08 | Ct = 31.6 ± 1.6 |
| SYND4 | F | TGAAGTTGTCCATCCCTTGGTG | R | TCAACGGGTGAGATTCTCTTGG | E = 1.95 ± 0.13 | Ct = 26.7 ± 3.0 |
| MuRF1/TRIM63 | F | TGTGCAGACCATCATCACTCAG | R | AACTTCTGGCTCAGCTCTTCC | E = 1.97 ± 0.04 | Ct = 25.9 ± 1.4 |
| MuRF2/TRIM55 | F | TGCCCATCCTGTAGACATGAAG | R | GCTGGTCGGATTTCTTTTCTGG | E = 1.98 ± 0.05 | Ct = 27.4 ± 1.1 |
| REDD1 | F | TGGACAGCAGCAACAGTGG | R | TCACTGAGCAGCTCGAAGTC | E = 1.93 ± 0.10 | Ct = 28.9 ± 2.9 |
| REDD2 | F | TTGCTGGACTGTGGCTATCAC | R | CAAGGACCTTTGAGCAACCAAG | E = 1.96 ± 0.10 | Ct = 26.5 ± 1.8 |
| SRF | F | CAGTGCAGGCCATTCAAGTG | R | AGTTGTGGGCACGGATGAC | E = 1.95 ± 0.09 | Ct = 28.0 ± 2.7 |
| Arkadia/RNF111 | F | GGCTATGGATCAAGCATGGTTG | R | GCTTGATGATGAAGTGGCCTTG | E = 1.84 ± 0.04 | Ct = 33.3 ± 1.5 |
| Atrogin-1/FBXO32 | F | CGATGTTACCCAAGGAAAGAGC | R | TCAGTGCCCTTCCAGGAAAG | E = 1.98 ± 0.13 | Ct = 25.3 ± 3.3 |
| cSKI | F | CGACGTGAAGGAGAAATTCGAC | R | CGGCTTGTCCTTTTCGGAAG | E = 1.95 ± 0.09 | Ct = 29.7 ± 2.9 |
| SnoN | F | GGGGCTTTGAATCAGCTAAATGG | R | TCCTGATGGTGCATCTGTCTTG | E = 1.89 ± 0.09 | Ct = 27.9 ± 2.8 |
| Titin | F | TGAGTTCCAGCAGCTTTATGGG | R | TAGGCGGAATTCCTCTTATGCC | E = 1.93 ± 0.05 | Ct = 20.6 ± 1.7 |
| LMNA | F | GAACATCTACAGTGAGGAGCTG | R | TCTCAAACTCACGCTGCTTC | E = 1.86 ± 0.04 | Ct = 30.9 ± 1.1 |
Values are expressed as means ± SD. Priming efficiencies (E) and cycle threshold (Ct) values are given in the two rightmost columns as an average of all quantitative RT-PCR reactions.
F, forward primer; R, reverse primer; PPIA, peptidylprolyl isomerase A; β2-m, b2-microglobulin; RPL32, ribosomal protein L32; PolR2A, polymerase (RNA) II (DNA directed) polypeptide A; MyHC1, 2A, and 2X, myosin heavy chain 1, 2A and 2X; PGC1α s4, peroxisome proliferator-activated receptor gamma, coactivator 1 alpha, splice variant 4; Ankrd2, ankyrin repeat domain 2; FGF6, fibroblast growth factor 6; IGF1Ea, insulin-like growth factor 1, splice Ea; PAX7, paired box 7; MCAD, M-cadherin; ITGA7, integrin, α7; CAV1, caveolin 1; MYOG, myogenin/myogenic factor 4; MYOD, myogenic differentiation 1/myogenic factor 3; MRF4/MYF6, muscle regulatory factor 4/myogenic factor 6; MYF5, myogenic factor 5; SYND4, syndecan 4; MuRF1/TRIM63, muscle-specific RING finger protein 1/tripartite motif containing 63, E3 ubiquitin protein ligase; MuRF2/TRIM55, muscle-specific RING finger protein 2/tripartite motif containing 55; REDD1 and 2, protein regulated in development and DNA damage response 1 and 2; SRF, serum response factor; RNF111, ring finger protein 111; FBXO32, F-box protein 32; cSKI, V-Ski avian sarcoma viral oncogene homolog; SnoN, SKI-like oncogene; LMNA, lamin A/C.