Fig. 3.

Gap19 reduces the expression of proinflammatory cytokines and the activation of astrocytes after ICH injury. a, b WB analysis showed the expression levels of TNF-α, IL-1β, IL-6, MCP1, and β-tubulin as the loading control. c, d The expression levels of IL-4 and IL-10. β-Tubulin was used as a loading control. e, f The EtBr uptake assay was used to evaluate the activity of Hcs on astrocytes, microglia, and neurons after hemin stimulation. Gap19 inhibits the hemin-induced excessive opening of Hcs on astrocytes. g, h Immunofluorescence for GFAP was used to evaluate astrocyte activation. TAT-Gap19 treatment significantly reduced the number of GFAP-positive cells in the area surrounding cerebral hemorrhage. i, j In vitro, Gap19 treatment reduced the GFAP fluorescence intensity in astrocytes after hemin stimulation. The bars in a–d, g, and h represent the SEM of the data from 3 cerebral hemorrhage tissue samples per group. *P < 0.05, **P < 0.01 compared to the sham group. #P < 0.05, ##P < 0.01 compared with the ICH group. The bars in e, f, i, j in the ICH group represent the SEM of data from 3 samples per group. *P < 0.05, **P < 0.01 compared to the control group. #P < 0.05, ##P < 0.01 compared with the hemin group