Fig. 7: Estrogen regulates the expression of immunosuppressive genes in the liver.

FC was performed on immune cells isolated 7 days post intrasplenic/portal injection of 5 × 10⁵ MC-38 cells and immunostained with the indicated antibodies. Shown in a (left) are results of an IFN-γ production assay performed on liver-derived CD8⁺ T cells extracted from OVX or sham-operated mice and in a (right) the mean proportions (%) of CD8⁺IFN-γ⁺ cells per liver (±s.e.m.) based on a pool of five livers per group, analyzed in triplicates. Shown in b are results of qRT-PCR (±s.d.) performed on RNA extracted from CD8⁺ T cells sorted from livers of tumor-injected mice (data normalized to GAPDH; n = 3). Shown in c are results of qRT-PCR performed on whole liver RNA obtained from tumor-bearing sham or OVX mice. The results for each of the indicated transcripts are based on livers obtained from three mice and expressed as means (±s.d.) relative to sham operated mice that were assigned a value of 1, all normalized to GAPDH. Shown in d, e are representative flow cytometric contour plots obtained with each of the indicated immune cells populations (left) and in the bar graphs (right) the relative frequency of TNFR2⁺ cells per liver, pooled from five livers per group and analyzed in duplicates. Shown in f are mean fluorescence intensity (MFI) plots (for TNFR2) obtained for the same CD4⁺CD25⁺Foxp3⁺ cells analyzed in e. Shown in g are results of qRT-PCR expressed as means (±s.d.) performed on RNA extracted from liver-derived CD4⁺Foxp3⁺ T cells isolated by cell sorting (data normalized to GAPDH; n = 3). *–p ≤ 0.05; **–p ≤ 0.01; ***–p≤ 0.001; NS, not significant as determined by the Student's t-test. Box and whiskers graphs: the box extends from the 25th to 75th percentiles, the middle line denotes the median and the whiskers extends from the minimum to the maximum value.