Quantification of Arg1 Expression in Murine Muscle

Real-time PCR was performed as per our published method [11 (link), 12 (link)]. Total RNA was isolated from the quadriceps muscles of mice. The muscle was homogenized and dissolved in TRIzol. RNA was isolated using the TRIzol method following the manufacturer's instructions, and the quality of the RNA preparations was monitored by absorbance at 260 and 280 nm (Helios Gamma, Thermo Spectronic, Rochester, NY). The RNA was reverse-transcribed into complementary deoxyribonucleic acid (cDNA) using iScript reagents from Bio-Rad on a programmable thermal cycler (PCR Sprint, Thermo Electron, Milford, MA). The cDNA (50 ng) was amplified by real-time PCR using a Bio-Rad iCycler and ABgene reagents (Fisher Scientific, Pittsburgh, PA) and ARG1 primers [12 (link)]. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal control for normalization.