Table 1. Primer sequences for real-time PCR.
Gene | Forward primer (5’ →3’) | Reverse primer (5’ → 3’) |
---|---|---|
Hypertrophy and proteolysis | ||
MGF | CGAAGTCTCAGAGAAGGAAAGG | ACAGGTAACTCGTGCAGAGC |
MSTN | GACCAGGAGAAGATGGGCTGAATCCGTT | CTCATCACAGTCAAGACCAAAATCCCTT |
MuRF-1 | GCCTTCTTCGCCTTCTCC | AGCTCATACAGACTCAGTTCC |
Atrogin-1 | ATGTGCGTGTATCGGATGG | AAGGCAGGTCAGTGAAGC |
Ribosome Biogenesis | ||
c-Myc | ACAACCGAAAATGCACCAGC | CGTTGTGTGTTCGCCTCTTG |
TAF1B | TCTTGAAAGTGGAGCGGAGT | GTGTCTGTGGCATGGTCATC |
45S pre-rRNA | GAACGGTGGTGTGTCGTT | GCGTCTCGTCTCGTCTCACT |
28S | GCGTTGGATTGTTCACCCAC | ACCTGTCTCACGACGGTCTA |
Exportin-5 | TGCCCTTCCATGAATGACCC | CTGAGTGGACCTTGAGGCTG |
Inflammation | ||
IL-6 | AGGAGACTTGCCTGGTGAAA | CAGGGGTGGTTATTGCATCT |
IL-10 | GAAGAGAAACCAGGGAGCCC | ATCCCTCCGAGACACTGGAA |
IL-1β | AAGGCGGCCAGGATATAACT | CCCTAGGGATTGAGTCCACA |
MCP-1 | TCCCAAAGAAGCTGTGATCTTCA | CAGATTCTTGGGTGGAGTGA |
TNF-α | TCCTTCAGACACCCTCAACC | AGGCCCCAGTTTGAATTCTT |
Satellite cell-related genes | ||
MyoD | AAGCGCCATCTCTTGAGGTA | GCGAGAAACGTGAACCTAGC |
Myogenin | GCCAGACTATCCCCTTCCTC | GAGGCCGCGTTATGATAAAA |
Pax7 | AGGGCCTCCTGCTTGTTTAT | GGTTTTGCCCAACTCAGTGT |
MHCemb | GAGGAGGCTGATGAACAAGC | TCCTGCTGGAGGTGAAGTCT |
Housekeeping genes | ||
Fbl | CCCACACCTTCCTGCGTAAT | GCTGAGGCTGTGGAGTCAAT |
GAPDH | AACCTGCCAAATATGATGAC | TCATACCAGGAAATGAGCTT |
All primers were designed using PrimerPlus3 (Cambridge, MA, USA) and BLASTed against other potential mRNA targets using the online NCBI Nucleotide database (Bethesda, MD). MGF, mechano-growth factor; mstn, myostatin; murf-1, muscle RING finger 1; TAF1B, TATA-box protein associated factor, RNA polymerase 1 subunit B; 45S pre-rRNA, 45S pre-ribosomal 5; IL, interleukin; MCP-1, monocyte chemoattractant protein-1; TNF-α, tumor necrosis factor α; MyoD, myogenic differentiation 1; Pax7, paired box 7; MHCemb, myosin heavy chain embryonic; fbl, fibrillarin; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.