Fig. 5

SRGN amplifies post-stroke neuroinflammation in vivo. A The qPCR analysis of Il1b, Tnf and Il6 mRNA levels of mice brain tissues 1 day after MCAO, injected with rSRGN or control solvent. B Representative immunofluorescence images showing the morphology of microglia (Iba1⁺, green) in the ipsilateral hemisphere of ischemia mice brains 1 day after MCAO, injected with rSRGN (2.5 mg/ mL) or control solvent. n = 4 mice per group. Scale bar, 100 μm. C, D Quantification of the average branches (C) and process length (D) of microglia based on the images in B. E, F Flow cytometry strategy labeling microglial IL-1β (E) and the quantification of IL-1β MFI (F) in the ischemic mice brains 1 day after MCAO, injected with rSRGN (2.5 mg/ mL) or control solvent. n = 3 ~ 4 per group. G The qPCR analysis of Il1b and Tnf mRNA levels of brain tissue from Srgn-KO mice and their WT littermates 1 day after MCAO. H, I Representative immunoblot images (H) and quantification (I) of the protein level of IL-1β, TNF-α and SRGN from brain tissue of Srgn-KO mice and their WT littermates 1 day after MCAO. GAPDH served as the loading control. n = 4 mice per group. J Representative immunofluorescence images showing the morphology of microglia (Iba1⁺, green) from Srgn-KO mice and WT counterparts 1 day after MCAO. n = 4 mice per group. Scale bar, 100 μm. K, L Quantification of the average branches (K) and process length (K) of microglia based on the images in J. Data represented as mean ± SEM, * p < 0.05, ** p < 0.01