Biometry of non-transgenic and gh-transgenic zebrafish (Danio rerio) of F0104 lineage. Total length (a) and total mass (b) in non-transgenic zebrafish and in gh-transgenic zebrafish. Data are expressed as mean ± standard error of the mean. Asterisk indicate significant difference mean values between groups (n = 21; p < 0.05).

Consensus stability analyses of the candidates reference genes (RGs) grouping non-transgenic and gh-transgenic groups of zebrafish (Danio rerio) of F0104 lineage. Final consensus stability analysis between all methods calculated by RefFinder algorithm (a). Consensus stability analysis of the candidates RGs grouping both the groups calculed by comparative delta Ct (dCt) method (b); and geNorm (c); NormFinder (d); and BestKeeper (e) algorithms.

Consensus stability analysis in non-transgenic and gh-transgenic zebrafish (Danio rerio) of F0104 lineage calculated by RefFinder algorithm. General gene expression stability in non-transgenic zebrafish (a) and in gh-transgenic zebrafish (b). Data are expressed as geometric mean of ranking values. The most stable genes are displayed on the left, and the least stable genes are displayed on the right of the X-axis.

Graphical representation of the pairwise variation analysis. Y-axis represents V value and X-axis represents the number of reference genes used to normalize the reaction. The lowest value is found for 2/3 reference genes, what means that the addition of a third reference gene is superfluous to obtaining reliable results in experiments using non-transgenic and gh-transgenic zebrafish (Danio rerio) of F0104 lineage.

Verification of the suitability of the reaction when normalized with the candidate reference genes ranked as the best for brain tissue (a,b) and for all tissues grouped (c,d), using brain gene expression results. As target genes, it was chosen the worst (a) and the third best (b) for brain. In all the graphics, the Y-axis represents mRNA relative expression, and X-axis indicates the experimental group: non-transgenic and gh-transgenic zebrafish of F0104 lineage. Data are expressed as mean ± standard error of the mean. Asterisk indicate significant difference mean values between groups (n = 21; p < 0.05).

Verification of the suitability of the reaction when normalized with the candidate reference genes ranked as the best for muscle tissue (a,b) and for all tissues grouped (c,d), using muscle gene expression results. As target genes, it was chosen the worst (a) and the third best (b) for muscle. In all the graphics, the Y-axis represents mRNA relative expression, and X-axis indicates the experimental group: non-transgenic and gh-transgenic zebrafish of F0104 lineage. Data are expressed as mean ± standard error of the mean. Asterisk indicate significant difference mean values between groups (n = 7; p < 0.05).

Verification of the suitability of the reaction when normalized with the candidate reference genes ranked as the best for intestine tissue (a,b) and for all tissues grouped (c,d), using intestine gene expression results. As target genes, it was chosen the worst (a) and the third best (b) for intestine. In all the graphics, the Y-axis represents mRNA relative expression, and X-axis indicates the experimental group: non-transgenic and gh-transgenic zebrafish of F0104 lineage. Data are expressed as mean ± standard error of the mean. Asterisk indicate significant difference mean values between groups (n = 7; p < 0.05).

Verification of the suitability of the reaction when normalized with the candidate reference genes ranked as the best for all tissues grouped (a,b), using liver gene expression results. As target genes, it was chosen the worst (a) and the second best (b) for liver. In the two graphics, the Y-axis represents mRNA relative expression, and X-axis indicates the experimental group: non-transgenic and gh-transgenic zebrafish of F0104 lineage. Data are expressed as mean ± standard error of the mean. Asterisk indicate significant difference mean values between groups (n = 7; p < 0.05).