Fig. 3

Effects of PD-1/PD-L1 overexpression and knockdown on brain cell death and neuronal degeneration after ICH. a ICH rats accepted intracerebroventricular injection of pDNA or siRNAs as indicated. b Western blot analysis of the efficiency of PD-1 and PD-L1 overexpression or knockdown in brain of ICH rats. Quantification of relative protein levels of PD-1 and PD-L1 was shown in c. c Data are expressed as mean ± SEM, n = 6. Triple asterisks indicate p < 0.001 vs. ICH + vector group; triple pound signs indicate p < 0.001 vs. ICH + control siRNA group. d Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Sections were labeled by TUNEL (green) to detect apoptotic brain cells and counterstained with DAPI (blue) to detect nuclei. Arrows point to TUNEL-positive cells. Scale bar = 64 μm. e Percentage of TUNEL-positive cells. Data are expressed as mean ± SEM, n = 6. Triple asterisks indicate p < 0.001 vs. sham group; double pound signs indicate p < 0.01 vs. ICH + vector group; ampersand indicates p < 0.05, double ampersands indicate p < 0.01 vs. ICH + control siRNA group. f Fluoro-jade B (FJB) staining. Arrows point to FJB-positive cells. Scale bar = 64 μm. The number of FJB-positive brain cells was calculated. g Data are expressed as mean ± SEM, n = 6. Triple asterisks indicate p < 0.001 vs. sham group; double pound signs indicate p < 0.01 vs. ICH + vector group; double ampersands indicate p < 0.01, triple ampersands indicate p < 0.001 vs. ICH + control siRNA group. h, i ELISA assay of the contents of IL-1β and TNF-α in the serum. The mean values of the sham group were normalized to 1.0. Data are expressed as mean ± SEM, n = 6. Single asterisk indicates p < 0.05 vs. sham group; Single pound sign indicates p < 0.05 vs. ICH + vector group; Single ampersand indicates p < 0.05, double ampersands indicate p < 0.01 vs. ICH + control siRNA group