Figure 5. Validating expression of enzymes involved in the biosynthesis of anabolic intermediates to confirm genes that were identified as differentially expressed in the maSigPro clustering analysis.

Porcine Longissimus Dorsi muscle transcript abundance of the following genes was determined by quantitative-RT-PCR to establish the effects of a beta-adrenergic agonist (grey bars; BA) or recombinant growth hormone (black bars; GH) treatment for 1–27 days, compared to a control cohort (white bars; C); Phosphoglycerate Dehydrogenase: Phgdh (A), Phosphoserine aminotransferase: Psat1 (B), Phosphoserine Phosphatase: Psph (C), the mitochondrial isoform of phosphoenolpyruvate carboxykinase: Pck2 (D), the cytosolic isoform of phosphoenolpyruvate carboxykinase: Pck1 (E), the muscle isoform of Pyruvate Kinase variant 1: Pkm1 (L), and Pyruvate Kinase variant 2, Pkm2 (M). Representative western blots (F,G) and quantification (H,I) of beta-adrenergic agonist (BA) and growth hormone (GH) treatment effects on Longissimus Dorsi (LD) muscle protein expression of Phosphoglycerate Dehydrogenase: PHGDH, and mitochondrial isoform of phosphoenolpyruvate carboxykinase: PEPCK-M, following 3 (F,H) and 7 (G,I) days of treatment. Alpha-tubulin was used as a loading control. Relationship between PHGDH and PEPCK-M protein expression in porcine Longissimus Dorsi muscle following treatment with a beta-adrenergic agonist (grey circles), growth hormone (black circles) or no treatment (white circles) following 3 (J) and 7 (K) days. Data is mean ± SEM. n = 10 for days 1, 3, 7 and 13, whilst n = 15 for day 27. *Indicates a treatment effect with p < 0.001 (unless otherwise stated). ^#Indicates a treatment x time interaction with p < 0.001. a Indicates a treatment effect with p < 0.01. ^§Indicates a treatment effect with p < 0.05.