Role of prolactin-like protein (PRL-L) in cold-induced increase of muscle mass in chicks

Clenbuterol changes phosphorylated FOXO1 localization and decreases protein degradation in the sartorius muscle of neonatal chicks

“…Real-time PCR was performed as described previously. 30) In brief, cDNA was synthesized from 40 ng of RNA per 10 μL of reaction solution using the PrimeScript RT Reagent Kit (RR036A; Takara, Shiga, Japan). Samples were incubated at 37°C for 15 min, 85°C for 5 s, and 4°C for 5 min.…”

“…Western blot analysis was performed as described previously. 30) In brief, samples were electrophoresed on a SDS/10% (w/v) polyacrylamide gel and then transferred to a polyvinylidene difluoride membrane (IPVH00010; Millipore Co., Billerica, MA, USA). The membrane was blocked with 5% (w/v) skim milk in Tris-buffered saline containing 0.1% (v/v) Tween-20 (TBST) for 1 h at room temperature.…”

Gene expression pattern of glucose transporters in the skeletal muscles of newly hatched chicks

“…The total RNA extraction with ISOGEN II (Nippon Gene, Tokyo, Japan) and reverse transcription reaction were performed as described in a previous study. 19) Realtime PCR was performed using the 7300 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA) with SYBR Select Master Mix (Applied Biosystems, Foster City, CA, USA). In this study, the following primers were used: GLUT1 (5′-GATGGCTTTGTCCTTTGAGATGC-3′ and 5′-CAAAGATGCTGGTGGAGTAGTAG-3′), GL-UT3 (5′-TTAGCTGGGTGTCGGTGCTGG-3′ and 5′-CCATGAGAGCACCCCCAGCAA-3′), GLUT8 (5′-CC-AAATGGGAACAACTCATCAA-3′ and 5′-GGGCAA-AACCAGCAACAAA -3′), GLUT12 (5′-TCTCTCTG-GCCAGCCTACTT-3′ and 5′-TTCACTCAGGACGAGC-CAAG-3′), hexokinase 2 (HK2) (5′-AAGCTCTCTTCG-GATCTGCG-3′ and 5′-CTTGGTGAGGATCTCGTGGG-3′), peroxisome proliferative-activated receptor, gamma, and coactivator-1 alpha (PGC-1α) (5′-GCAATTGAA-GAGCGTCGTGT-3′ and 5′-CCATAGCTGTCTCCAT-CATC-3′), insulin receptor (INSR) (5′-GACTCTCCA-ACGAACAGGTG-3′ and 5′-TCAGCATCTCAATGAC-CTCAA-3′), Insulin-like growth factor-I (IGF-I) (5′-CTTCAGTTCGTATGTGGAGACA-3′ and 5′-GATT-TAGGTGGCTTTATTGGAG-3′), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (5′-CCTCTCTG-GCAAAGTCCAAG-3′ and 5′-CATCTGCCCATTT-GATGTTG-3′).…”