Fig 2: The influence of anti-BTLA agonist antibody treatment on cellular and molecular neuroinflammation following acute ICH. A Schematic illustrating selected regions from three comparable sections per mouse for quantitative analysis of Iba-1+, GFAP+, MPO+, and FJB+ cells in the perihematomal regions. B Representative immunofluorescence images of Iba-1 (microglia/macrophages), GFAP (astrocytes), and MPO (neutrophils), along with FJB staining for degenerating neurons in perihematomal regions at day 3 post-ICH. Insets show magnified views of positive cells (scale bar = 50 μm). C Scatter plots quantifying Iba-1+, GFAP+, MPO+, and FJB+ cell densities. D Representative Western blot bands of IL-1β, IFN-γ, RANTES, and HMGB1 expression in tissue surrounding the hematoma at 36 h post-ICH. β-actin was used as the loading control. E Quantification of IL-1β, IFN-γ, RANTES, and HMGB1 expression in the hemorrhagic brain 36 h after ICH in sham-operated and ICH mice treated with isotype control or anti-BTLA agonist. Bands used for statistical analysis were those exposed individually, not the re-exposed bands from the reassembled membranes provided in the supplementary materials. *P < 0.05, **P < 0.01. All data are presented as mean±SD or median (interquartile range)

Fig 3: The influence of microglia depletion on the effects of an anti-BTLA agonist for molecular neuroinflammation in mice with acute ICH. A Double fluorescent staining of Iba-1 (green) and BTLA (red) in the perihematomal region 3 days after ICH. Nuclei were counterstained with DAPI (blue). Merged images are shown (scale bar = 50 μm). B Representative immunofluorescence images showing Iba-1+ cells in the perihematomal region of microglia-depleted mice 3 days post-ICH (scale bar = 50 μm). C Quantitative analysis of Iba-1+ cells in mice that received vehicle or CSF1R inhibitor, which was used for the depletion of microglia on day 3 after ICH. D Representative Western blot bands of IL-1β, IFN-γ, RANTES, and HMGB1 expression in tissue surrounding the hematoma at 36 h post-ICH. E Quantitative analysis of IL-1β, IFN-γ, RANTES, and HMGB1 expression in ICH mice that received vehicle or CSF1R inhibitor. Statistical analysis was performed on individually exposed bands, not on the re-exposed versions presented in the reassembled full-membrane supplementary figures. **P < 0.01. All data are presented as mean±SD