Fig 1: Sex Hormones and the GH Signaling Network Are Perturbed in GF Mice(A and B) Serum levels of testosterone (A) and E2 (B) in male and female ConvR and GF mice.(C) Circulating GH total secretion, peak number, and levels in male and female ConvR and GF mice.(D) Representative examples of temporal profiles of GH secretion in ConvR♂, GF♂, ConvR♀, and GF♀ animals.(E) Enrichment analysis of FOXA1, FOXA2, BCL6, CUX2, and STAT5 (male and female specific) target genes for differentially expressed genes of indicated contrasts (♂ = GF♂ versus ConvR♂; ♀ = GF♀ versus ConvR♀; ConvR = ConvR♂ versus ConvR♀; GF = GF♂ versus GF♀). Target genes have been identified by published chromatin immunoprecipitation sequencing (ChIP-seq) data.(F) Phosphorylation of STAT5 (Tyr694) in male and female ConvR and GF mice.(G) Hepatic mRNA and protein levels of the other GH-dependent TFs BCL6 and CUX2 that are involved in sex-dimorphic gene expression for the indicated conditions.(H) Experimental design to mimic male-specific GH pulses in GF mice.(I) Volcano plot of expression difference in mRNA expression between GH-injected and vehicle-injected GF male mice show a rescue in sex-biased gene expression.(J) Enrichment analysis of male- and female-specific genes (♂ biased, ♀ biased) or repressed or induced genes in GF mice (down or up in GF♂) for differentially expressed genes of the contrast GH- versus vehicle-injected GF♂ mice.(K) Experimental design of an in vitro system of primary hepatocytes treated with sera of different sources (ConvR♂, ConvR♀, and GF♀).(L) Volcano plot of expression difference in mRNA expression of female hepatocytes treated with GF♀ serum versus ConvR♀ serum. Color code corresponds to (B).(M) Enrichment analysis of male- and female-specific genes (♂ biased, ♀ biased) for differentially expressed genes of the indicated contrasts (serum treatment: ConvR♂ versus ConvR♀ or GF♀ versus ConvR♀) in female hepatocytes.(N) Cartoon depicting the experimental design to evaluate the effect of a tonic GH treatment.(O) Number of differentially expressed genes in function of an adjusted p value threshold for the indicated contrast. GH treatment diminished the number of differentially expressed genes between female hepatocytes treated with GF♀ serum and ConvR♀ serum.(P) Enrichment analysis of male- and female-specific genes (♂ biased, ♀ biased) for differentially expressed genes between GH-treated versus untreated female hepatocytes.All boxplots represent at least 12 independent biological replicates per condition, except for (G), for which at least six biological replicates were used. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001. All boxplots are Tukey boxplots.