Fig. 5: Estrogen promotes an immunosuppressive microenvironment 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 the contour plots obtained for each group and in a (right) the proportion (%) of CD11b⁺Ly6C⁺Ly6G⁺ cells per total liver immune cells expressed as means (±s.d.) of three experiments, each performed on a pool of five livers per group. Shown in b, c and e are representative confocal images from 10 μm cryostat liver sections obtained 7 days post intrasplenic/portal inoculation of 5 × 10⁵ MC-38 cells and immunostained with the indicated antibodies followed by Alexa Fluor 568 (green) for Gr-1 (b), Alexa Fluor 647 (red) for CD8 (d), Alexa Fluor 647 (green) for F4/80 (e), and 4′,6-diamidino-2-phenylindole (DAPI; blue in b and c, and red in e). Shown on the right of the confocal images are the numbers (means ± s.e.m.) of the indicated cells per field counted in 7 fields (OVX, OVX + E2) or 12 fields (sham) per group. Shown in d (left) are representative flow cytometric contour plots obtained for the indicated groups and in d (right) the mean proportion of CD11b⁺CD11c⁺ cells per liver based on a pool of five livers per group, analyzed in triplicates. *−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. Scale bars correspond to 100 µm on IF images.