Fig 2: Gross assessment at 24 h and 72 h. (A) SAH grade indicated that all mice suffered from severe (≥ 13) SAH, and no difference was found among the SAH groups even administered with GPNMB. Anyhow, (B) mNSS and (C) Garcia test showed that neuronal function was compromised significantly after SAH induction and could be rescued by using GPNMB. Similarly, (D) BWC and (E) BBB integrity analyses showed that brain edema and BBB damage were significant after SAH, and administration of GPNMB had a protective effect especially at medium and high dose; (F) SAH grade results suggested the severe SAH, but no significances were noted among the vehicle and GPNMB groups. Results from (G) mNSS and (H) Garcia test showed that neurofunction was significantly impaired after SAH, and it was improved with GPNMB use. Additionally, (I) BWC and (J) BBB integrity had a significant defect after SAH, and were to some extent protected by using GPNMB, ANOVA, *p < 0.05 vs. sham group, #p < 0.05 vs. vehicle group, n = 6 / group

Fig 3: Immunofluorescence (IF) staining for GPNMB. GPNMB (green) costaining with A microglia (red), B astrocytes (red) and C neurons (red), from it we could see that GPNMB was expressed abundantly in microglia, astrocytes and neurons; all fields of vision were taken on the left cortical and subcortical areas, n = 2 / group

Fig 4: Enzyme linked immunosorbent assay (ELISA) results. ELISA was utilized to additionally confirm the expression of A IL-1β, B IL-6 and C TNF-α, and the results were consistent with that from WB that the expression of the inflammatory cytokines surged after SAH and could be decreased with GPNMB, ANOVA, *p < 0.05 vs. sham group, #p < 0.05 vs. vehicle group, @p < 0.05 vs. GPNMB group, n = 6 / group

Fig 5: Mid- and long-term neurofunction tests. Rotarod test at (A) 5 RPM and (B) 10 RPM suggested that stable duration was significantly decreased after SAH and increased with the administration of GPNMB. Morris water maze test showed that (C) distance traveled and (D) escape latency was significantly dropped after SAH while had an increase with GPNMB usage. Also, (E) retaining time in the target quadrant was improved significantly in GPNMB group, and this was consistent with the (F) heatmap demonstration, ANOVA, *p < 0.05 vs. sham group, #p < 0.05 vs. vehicle group, n = 6 / group