Fig. 3: Splenectomy reduces cardiac infiltration of monocytes and heart failure in 5/6Nx mice.

a Induction of GPR68 expression in circulating monocytes in 5/6Nx mice. Flow cytometry analysis was performed to detect high-GPR68-expressing CD11b⁺/Ly6C⁺ cells in the spleen, blood, and bone marrow of Sham and 5/6Nx mice. b Decrease in the number of cardiac-infiltrated CD11b⁺/Ly6C⁺ cells in splenectomized (Splx)-5/6Nx mice. Left panel shows the flow cytometry histogram of CD11b⁺/Ly6C⁺ cells isolated from the cardiac ventricle of Sham, 5/6Nx, Splx, and Splx-5/6Nx mice. The number of CD11b⁺/Ly6C⁺ cells, indicated by red circles, was quantified and the data are shown in the right panel. c Serum concentrations of BNP in Sham, 5/6Nx, Splx, and Splx-5/6Nx mice. d The mRNA levels of inflammatory cytokines (Tnf-α and IL-6) and fibrosis-related factors (αSma and Col1a2) in the cardiac ventricle of Sham, 5/6Nx, Splx, and Splx-5/6Nx mice. The mean value of the sham-operated wild-type group was set as 1.0. e Double immunofluorescence labeling of GPR68 with F4/80 in the ventricle slices prepared from Sham, 5/6Nx, Splx, and Splx-5/6Nx mice. High-GPR68-expressing cells are double-labeled (yellow) with F4/80. The scale bar indicates 50 μm. f Decrease in the number of cardiac-infiltrated high-GPR68-expressing monocytes in Splx-5/6Nx mice. For all panels, graphs show the mean ± SD of individual mice in independent experiments. Statistical significance was determined using two-way ANOVA with Tukey–Kramer post hoc tests (b–d, f). Numbers and P-values are shown in each graph. Source data are provided as a Source Data file.