Table 1

Primer sequences for RT-PCR"

Table 2

Primary and secondary antibodies used in Western blot"

Fig. 1

Effects of MET on chicken growth and growth-related hormones A. Effect of MET on chicken growth (ADFI. average daily feed intake; AGD. average daily gain; F/G ratio. feed/gain ratio; Data are expressed as "$\bar x \pm s$" (n=4, 40 chicks per group), *. P < 0.05 compared with the control group, according to the t test); B. Serum ATP level; C. Serum insulin (INS) level; D. Serum levels of growth hormone (GH), insulin-like growth factor 1 (IGF1), and insulin-like growth factor binding protein 2 (IGFBP2); in Fig. B-D, data are represented as "$\bar x \pm s$" (n=6 per group), *.P < 0.05 for the female treatment group compared with the female control group, **.P < 0.01 for the female treatment group compared with the female control group; #. P < 0.05 for the male treatment group compared with the male control group, ##. P < 0.01 for the male treatment group compared with the male control group, according to the t test"

Fig. 2

Effects of MET on ATP level and mitochondrial respiratory enzymes in the liver, kidney, and muscle of chicks A. ATP level; B. Nicotinamide adenine dinucleotide hydrogen (NADH) concentration; C. Cytochrome C oxidase activity; D. ATP synthase activity; Data are represented as "$\bar x \pm s$" (n=6 per group), *. P < 0.05 for the female treatment group compared with the female control group, **. P < 0.01 for the female treatment group compared with the female control group; #. P < 0.05 for the male treatment group compared with the male control group, ##. P < 0.01 for the male treatment group compared with the male control group, according to the t test"

Fig. 3

Effects of MET on mRNA levels of genes related to growth metabolism in the liver, kidney, and muscle of chicks A. Relative mRNA levels of ATP synthase F1 subunit α (ATP5A1), liver kinase B1 (LKB1), AMP-activated protein kinase α2 (AMPKα2), and mammalian target of rapamycin (mTOR) in the liver; B. Relative mRNA levels of insulin-like growth factor 1 (IGF1), insulin-like growth factor 1 receptor (IGF1R), insulin (INS), insulin receptor (INSR), phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), and AKT serine/threonine kinase 1 (AKT1) in the liver; C. Relative mRNA levels of p21, p27, Cyclin-dependent kinase 2 (CDK2), and Cyclin E1 in the liver; D-F and G-I show relative mRNA levels of the above genes in the kidney and muscle respectively; Data are represented as "$\bar x \pm s$"(n=3 per group), *. P < 0.05 for the female treatment group compared with the female control group, **. P < 0.01 for the female treatment group compared with the female control group; #. P < 0.05 for the male treatment group compared with the male control group, ##. P < 0.01 for the male treatment group compared with the male control group, according to the t test"

Fig. 4

Effects of MET on protein expressions related to growth metabolism in the muscle of chicks A. Western blot analysis of protein bands; B. p-LKB1/LKB1 ration; C. p-AMPKα2/AMPKα2 ration; D. p-mTOR/mTOR ration; E. Relative protein level of ATP5A; F. Relative protein levels of IGF1R, INSR, PIK3CA, and AKT1; G. Relative protein levels of p21, p27, CDK2, and Cyclin E1; Data are represented as "$\bar x \pm s$" (n=3 per group), *. P < 0.05 for the female treatment group compared with the female control group, **. P < 0.01 for the female treatment group compared with the female control group; #. P < 0.05 for the male treatment group compared with the male control group, ##. P < 0.01 for the male treatment group compared with the male control group, according to the t test"

Fig. 5

Mechanism diagram of MET regulating the growth metabolism of chicks through the LKB1/AMPKα2 pathway"