Figure 5.

HMGB2 promotes the transcription of Ctss. (A) patterns of Hmgb2 and Ctss transcripts in the cortex of mice at 3 days after operation with stroke, as compared to sham. (B) the lystaes of cytosolic fraction and extracellular space from mice at 1 or 7 days after operation with sham or stroke were blotted with anti-Ctss or α-tubulin, as indicated. C, Relative expression (R.E) levels (defined by normalizing the band intensities of anti-Ctss blots to the respective α-tubulin) in the individuals (circles) and their averages per group (triangles) are plotted. Data are mean ± SEM (n = 5 mice per group, ns = no significantly differences, ***p < 0.0001, t-tests). (D) illustration (top) shows a direct binding of Hmgb2 to a promoter region of Ctss. CHIP- qualificative PCR assay shows amplification of the predicted Hmgb2-binding sites in Ctss promoter precipitated with anti-Hmgb2 or non-specific IgG, Relative expresssion of each site in the individuals (circles) and their averages per group (triangles) are plotted (left), n=3, representative image shows the bands of P3 PCR product (right) . (E) The relative luciferase activity (units) in the indiviudal assay (circles), in which a full length (1425 bp)of Hmgb2 binding sites located in Ctss promoter or GFP was co-expressed with an empty vector or Hmgb2, and their averages per group (triangles) are plotted (bottom). Data are mean ± SEM (n = 5 assays per group, F_{(2, 12)} = 18.72, ns = no significantly differences, ***p < 0.0001, BF ANOVA).