FIG. 2.

Altered gene expression (A) and key regulatory proteins in growth signaling pathway (B) in adult female homozygous thrab 1-bp insertion (m/m) mutant fish. The mRNA expression of gh1, smtla, and smtlb in pituitary (A-I) and mRNA expression of igf-1a, igf-1b, igf-1ra, igf-1rb, Ir-a, glut4, and myo in the muscles (A-II) of WT (solid bars) and sibling homozygous thrab 1-bp insertion (m/m) mutant fish (open bars) were determined by RT-PCR as described in Materials and Methods section. Number of fish = 10–15 for pituitaries, 5–3 for muscles. (B-I) Western blot analysis was carried out for p-AKT (Ser473), total AKT, p-p70S6 (Thr389), total p70S6, p-S6 (Ser240/244), total S6, and GAPDH, using muscle as described in Materials and Methods section. (B-II) Quantitative analysis of relative protein expression levels of the ratios of p-AKT to total AKT (a), p-p70S6 to total p70S6 (b), and p-S6 to total S6 (c) using GAPDH as a loading control. The data are shown as mean ± SE (n = 3; the p-values are indicated). Two-tail unpaired t-test, p-adjusted <0.05 was used for statistical analysis. mRNA, messenger RNA; RT-PCR, real-time PCR.