Fig 2: Blood levels of Gal-9 and TIM-3-positive T cells are markedly increased in patients with AD. (A) Serum Gal-9 levels of patients with atopic dermatitis involved in this study (AD, n = 30), as compared to healthy controls (HC, n = 30), (B) the percentage of CD4+TIM-3+ T cells in whole blood, the percentage of (C) TIM-3+TH1 cells and (D) TIM-3+TH17 cells in PBMCs of the above patients with AD compared to HC. These results are presented as means ± SEM.

Fig 3: Tim-3 was increased in the renal macrophages of DN mice. Western blot (A) and RT-PCR (B) analyses showing the expressions of Tim-3 in kidneys from Ctrl, STZ and db/db mice. (C) Representative photomicrographs of Periodic Acid-Schiff (PAS) staining and immunohistochemical (IHC) staining of Tim-3, CD68, Nephrin, and WT-1 in renal glomeruli of different mice. (D) Representative photomicrographs of double-labeled immunofluorescence (IF) staining of Tim-3 (red), CD68 (green) and nucleus (blue) in renal glomeruli of STZ mice. Bar = 10 μm. (E) Representative flow cytometry (FCM) of Tim-3 expressions in renal macrophages of different mice. The bars represent the mean values and the stand, ad errors of the means (Mean ± SEM). *P < 0.05, **P < 0.01, ***P < 0.001 vs normal control.

Fig 4: Tim-3 accelerated podocyte injury depending on the NF-κB/TNF-α signaling pathway in macrophages. MPC were stimulated with different CM from PMs in the following experiments. (A–B) Western blot analysis showing the relative protein level of Nephrin in podocytes stimulated with different CM. (C–D) Confocal microscopy analysis showing the expression and quantification of actin cytoskeleton stimulated with different CM. *P < 0.05, **P < 0.01 vs controls. Data are expressed as means ± SEM, Student's t-test was employed for comparisons between two groups; one-way ANOVA followed by Tukey's post-test for multiple comparisons was used for groups of three or more.

Fig 5: Flow cytofluorimetric analysis workflow. The image shows the sequential procedure and the microphotographs of stained cells (A–H) using five different monoclonal fluorescent dye-coupled anti-human antibodies and analysed by flow cytometry (Amnis FlowSight). For each patient, four aliquots of lymphocyte suspensions (100 µL) were stained with antibodies using four different combinations (10 µL of each antibody). Representative stained cells (event) for each of the four combinations are shown: CD-3+ CD-4+ TIM-3+ (A); CD-3+ CD-4+ Dectin-1+ (B); CD-3+ CD-8+ TIM-3+ (C); CD-3+ CD-8+ Dectin-1+ (D). Tim3− and Dectin-1− cells are also shown: CD3+ CD-4+ TIM-3− (E); CD-3+ CD-4+ Dectin-1− (F); CD-3+ CD-8+ Tim3− (G); CD-3+ CD-8+ Dectin-1− (H).