Fig. 3: Warm temperature attenuates sexual dimorphism.

A The numbers of DEGs between male and female in different temperature-treated groups among multiple organs. B Warm temperature feminizes gene expression in males but masculinizes gene expression in females. Volcano plot of hepatic sex-biased genes in the Cool-acclimated group (left). Differences in hepatic gene expression between Warm-acclimated males and Cool-acclimated males categorized by sex-biased expression in Cool-acclimated frogs (middle). Differences in hepatic gene expression between Warm-acclimated females and Cool-acclimated females categorized by sex-biased expression in Cool-acclimated frogs (right). Colors indicate male-biased (red), female-biased (green), and unbiased (gray) expressed genes. The dotted lines paralleling to x axis or y axis indicate |log2 FC| = 1 or −log10 adjust p-value = −log10 0.05, respectively. CA Cool-acclimated, WA Warm-acclimated. C The top 5 hepatic KEGG pathways were significantly enriched between male and female in the Cool-acclimated group. And the enrichment analysis of these corresponding pathways between the indicated contrasts. *FDR adjusted p < 0.05, **<0.01, ***<0.001. (male and female in a specific temperature-treated group, left; males in different temperature-treated groups, middle; females in different temperature-treated groups, right; CA Cool-acclimated, CS Cool-switched, WS Warm-switched, WA Warm-acclimated). D Heatmap showing the hepatic expression of genes involved in the steroid biosynthesis pathway between temperature-switched groups and temperature-acclimated groups and significantly expressed at least one condition in male or female. Gene expression is shown as the z-score of FPKM (fragments per kilobase of transcript per million mapped reads). The numbers on the right indicate the log2 FC values between male and female in a specific temperature-treated group. Significant differential gene expression was detected in at least one condition. E Representative images of testis (unilateral) in each temperature treatment (scale bars, 1 mm). F The histogram indicates the weight and the line indicates the relative weight of the testis (testis weight versus body weight) in each temperature treatment. The data are given as mean ± SEM. G Venn plots show the shared DEGs between indicated contrasts across multiple organs in males and females. H Summary of the relationship between sex phenotype, intron retention, and ambient temperature across lizard, frog, and mouse. The data are presented as mean ± SEM. Colored lines are described as in Fig. 1. All experiments were conducted based on six biological replicates in each temperature-treated group with male:female = 3:3 (except Cool-switched group, which is 4:2).