Hierarchical clustering of the six liver transcriptomes and real-time RT-PCR validation of RNA expression of selected genes.

(a) Hierarchical clustering of the six liver transcriptomes from control males and females, E2-treated males and females, and KT11-treated males and females. The three female samples were closely clustered together, and E2-treated males was clustered with the female samples. Control males and KT11-treated males were clustered together and were distinct from the others. Scale bar represented the pearson correlation score. Heatmap was constructed with transcripts that showed significant differences in at least one comparison. (b-e) Real-time RT-PCR validation of transcripts enriched in the female liver (b), male liver (c), induced by KT11 (d) or E2 (e) in male livers. Fold changes (log2 base) measured by real-time RT-PCR (blue bars) are compared with those measured by RNA-SAGE sequencing (red bars).

Intersections of differentially expressed genes in the male zebrafish livers after E2 or KT11 treatment with sex-biased genes.

(a) Venn diagram of up-regulated genes in E2-treated males overlapped with sex-biased genes. (b) Venn diagram of down-regulated genes in E2-treated males overlapped with sex-biased genes. (c) Venn diagram of up-regulated genes in KT11-treated males overlapped with sex-biased genes. (d) Venn diagram of down-regulated genes in KT11-treated males overlapped with sex-biased genes . (e) Heat map of expression changes of sex-biased genes in E2- and KT11-treated male livers. For the column corresponding to the sex-biased transcripts, red represents female-biased genes and green represents male-biased genes. For the two columns corresponding to the E2- and KT11-treated male livers, red represents up-regulation and green represents down-regulation. The color intensity is calculated by logarithm-transformed (base 10) p-value. (f) Venn diagram of overlap of up-regulated genes in E2-treated male and down-regulated genes in KT11-treated males. (g) Venn diagram of overlap of down-regulated genes in E2-treated male and up-regulated genes in KT11-treated males. (h) Venn diagram of overlap of up-regulated genes in E2-treated male and up-regulated genes in KT11-treated males. (i) Venn diagram of overlap of down-regulated genes in E2-treated male and down-regulated genes in KT11-treated males.

Changes of sex-biased GO categories in E2- and KT11-treated male zebrafish liver.

Female-biased GO categories are in yellow shade and male-biased GO categories are in blue shade. GO categories under different classification (biological process, molecular function, and cellular component) are combined into functional categories to facilitate the interpretation. Red color represents up-regulation of the GO category and green color represents down-regulation of the GO category. It is clear that most of the female-biased GO categories were up-regulated in E2-treated males and down-regulated in KT11-treated males, while most of the male-biased GO categories were not affected.

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