Fig. 4

Depletion of PMN-MDSCs worsened brain injury, increasing microglial activation and inflammatory factor expression. PMN-MDSC depletion in hypoxia model of 7-day-old mice model to analysis (A–L): (A) Statistical analysis of the percentage of brain infarct area in Ly6G mAb group and IgG2b mAb group (n = 4). (B) Statistical analysis of brain water content results in Ly6G mAb group and IgG2b mAb group (n = 4). (C) Statistical analysis of geotaxis reflex time in Ly6G mAb group and IgG2b mAb group (n = 4). (D) Representative HE staining image (original magnification, ×400) in IgG and Ly6G antibody groups. (E) Representative Nissl staining image (original magnification, ×400) and (F) statistical analysis of the hippocampal neuronal damage scores in Ly6G mAb group and IgG2b mAb group (n = 5). (G) Representative images of NeuN (red) and TUNEL staining (green) in the hippocampal region and DAPI staining is shown in blue. Scale bar = 50 μm. (H) Statistical analysis of the percentage of TUNEL cells and the number of NeuN-TUNEL positive mouse hippocampal neurons in Ly6G mAb group and IgG2b mAb group (n = 4). (I) Representative merged and enlarged images showing CD68, IBA1, and DAPI co-staining in the hippocampus (Scale bar = 50 μm). (J) Statistical analysis of the number of CD68⁺ IBA1⁺ double positive microglia cells within the hippocampal for Ly6G mAb group and IgG2b mAb group (n = 4). (K) Statistical analysis of CD80/CD206 and CD86/CD206 relative ratios in Ly6G mAb group and IgG2b mAb group (n = 5). (L) Statistical analysis of p-STAT1, pNF-κB Thr254, and pNF-κB Ser529 levels in microglia from Ly6G mAb group and IgG2b mAb group (n = 5). Data are pooled from two independent experiments; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Bar graphs show the mean ± SEM. Statistical significance was determined using a two-tailed unpaired Student’s t-test (A- C, F, H, J - L)