Figure 3

High dietary salt intake enhances bone loss by modulating Th17-Treg cell balance. Cells from bone marrow (BM) of normal, low salt diet (LSD) and high salt diet mice groups (HSD) were isolated at the end of experiment, labelled and analysed by flow cytometry for percentage of Foxp3⁺ and Rorγt⁺, IL-10⁺Foxp3⁺ and Rorγt⁺IL-17⁺ cells. Gating was first performed for CD4⁺ and later analysed for the expression of CD4⁺Foxp3⁺ (Tregs), CD4⁺Rorγt⁺ (Th17), CD4⁺IL-10⁺ (Tregs) and CD4⁺IL-17⁺ (Th17). (A) Average percentage of CD4⁺Foxp3⁺T cells (B) Average percentage of CD8⁺Foxp3⁺T cells (C) Average percentage of CD4⁺Rorγt⁺T cells. (D) Average percentage of Foxp3⁺IL-10⁺ (Treg). (E) Average percentage of Rorγt⁺ IL-17⁺ (Th17). The results were evaluated by using ANOVA with subsequent comparisons by Student t test for paired or nonpaired data, as appropriate. Analysis was performed using Sigma plot software (Systat Software, Inc., Germany). Values are reported as mean ± SEM (n = 10) and similar results were obtained in three independent experiments. Statistical significance was defined as p ≤ 0.05 (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001) with respect to low salt diet group.