Fig. 7

Immunofluorescence staining of cleaved-caspase-3⁺ and CD3⁺ in the spleen samples from each group of mice. A Double immunofluorescence staining in the sham, CLP, anti-IL-6, anti-PD-1, and anti-IL-6/PD-1 mice groups were used to analyze the expression levels of cleaved-caspase-3⁺ and CD3⁺ in the spleen cells of mice. Cleaved-caspase-3⁺ cells and CD3⁺ cells showed red and green fluorescence, respectively. The nuclei were stained using DAPI (blue fluorescence). The final panel shows the merged double-positive CD3⁺ + cleaved-caspase-3⁺ + DAPI-stained cells. B Quantification of CD3-positive T-cell count per field. C Quantification of cleaved caspase-3-positive apoptotic cell count per field. Data are expressed as median and interquartile range; *p < 0.05, **p < 0.005, ***p < 0.001, as determined by one-way ANOVA with post-hoc Tukey's multiple comparison test. Scale bar: 50 μm