Fig 2: Representative staining images of the colon sections from UC patients and healthy donors. The arrows indicate eosinophils, CD177+, and CD40L+ cells, respectively. The scar bar represents 100 μm. (A–C) Characteristics of eosinophils, CD177+ and CD40L+ cells, respectively, in healthy donors. (D–F) Characteristics of eosinophils, CD177+, and CD40L+ cells, respectively, in UC patients who achieved histological remission. (G–I) Characteristics of eosinophils, CD177+, and CD40L+ cells, respectively, in UC patients who failed to achieve histological remission. A-UC: UC patients who stayed histological activation; HC: Healthy donors; R-UC: UC patients who achieved histological remission; UC: Ulcerative colitis.

Fig 3: Müller cells were left untreated or treated with soluble CD40L for 24 h. Levels of vascular endothelial growth factor (VEGF) (A), matrix metalloproteinase-9 (MMP-9) (B), monocyte chemotactic protein (MCP)-1 (C) and high-mobility group box-1 (HMGB1) (D) were quantified in the culture media by ELISA. Protein expression of CD40 (E), caspase-3 (F) and glial fibrillary acidic protein (GFAP) (G) in the cell lysates was determined by Western blot analysis. sCD40L stimulated proliferation of Müller glial cells that was almost as potent as 10 ng/mL of VEGF compared to untreated cells (H). Results are expressed as median (interquartile range) from three different experiments. (* p < 0.05 compared with the values obtained from untreated cells; Mann–Whitney test).

Fig 4: ZEB1 was poorly expressed in both postmenopausal osteoporotic patients and rats. A The heatmap analysis of differentially expressed genes in the GSE56815 dataset where BMD refers to bone mineral density. B Detection of ZEB1 expression in the blood of normal controls and OP patients by RT-qPCR (n = 40). C Detection of ALP, OPN and RUNX2 expression in bone tissues of sham- and OVX-treated rat by RT-qPCR. D Detection of TRAP expression in rat bone tissue by immunohistochemistry. E Assessment of bone content and structural changes in femoral tissues of rats by HE staining where bone trabeculae were labeled with red arrows and bone mass was labeled with green arrows. F–G Expression of ZEB1 in rat bone tissues at mRNA and protein levels by RT-qPCR (F) and Western blot (G). All data are represented as mean ± SD (n = 6 for rats in each group). Each experiment was performed three times independently. *p < 0.05, **p < 0.01. Differences are tested using two-way ANOVA with Tukey’s post hoc test (C) and unpaired t-test (B, D, F, G)

Fig 5: CD40 expression on human retinal microvascular endothelial cells (HRMECs) was evaluated through flow cytometry. Representative histogram of 4 independent experiments showing mean fluorescence intensity (MFI) of CD40 expression on HRMECs. CD40 expression on HRMECs stained with the primary mouse anti-human CD40 antibody and secondary goat anti-mouse AlexaFluor488 (blue) compared to cells incubated with the secondary antibody only (red) (A). HRMECs were left untreated (Control “C”) or treated with tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), TNF-α plus a 1 h-pre-incubation with BAY11-7085 or VEGF plus a 1 h-pre-incubation with BAY11-7085 for 24 h. Protein expression levels of CD40 (B,D–F) and CD40L (C) in cell lysate were determined by Western blot analysis. Results are expressed as mean ± standard deviation from three different experiments. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. (* p < 0.05 compared with the values obtained from untreated cells. # p < 0.05 compared with TNF-α-or VEGF-treated cells).