Fig. 6

Metformin reduced spleen weight and area of T- and B-cell zone of the spleen in rat recipients. a Macroscopic and microscopic (hematoxylin and eosin [HE], Gamma: 1.0; double-immunofluorescence [red: p-AMPKα, green: CD3, blue: nuclei; red: p-AMPKα, green: CD20, blue: nuclei]; objective magnification: 4 × or 20 ×; Gamma: 1.4) images of the spleen. Region accumulating with CD3⁺ or CD20⁺ cells represented T-cell zone or B-cell zone and marginal zone, respectively. The marginal zone additionally possesses erythrocytes (nonspecific double-stained structure without nuclei). b Boxplots represent the percentage of spleen weight/body weight. c Structures of white pulp and marginal zone of the spleen in rats. d Boxplots representing area percentage of T-cell zone, B-cell zone, and marginal zone. Boxplots represent the relative expression value of p-AMPKα per CD3⁺ T cells in the T-cell zone and CD20⁺ B cells in the B-cell zone. The relative expression value of p-AMPK was represented as mean fluorescence intensity (MFI) of p-AMPKα/ MFI of DAPI ratio in each field of view. A total of 200 cells from 10 HPF was measured for each rat. One-way analysis of variance (one-way ANOVA) followed by Tukey’s multiple test, *P, 0.05, **P, 0.01. (n = 6 rats/group)