Fig. 6

PMN-MDSCs from HIF-1α deficient mice exhibit reduced brain injury mitigation, with heightened microglia activation and inflammatory factor expression. Adoptive transfers experiment from 7-day-old mice in HIF-1α deficiency mice to analysis (A–L): (A) statistical analysis of the percentage of brain infarct area in adoptive transfer assay for PMN-MDSCs from hypoxia-treated HIF-1α^fl/fl mice and HIF-1α^fl/fl S100A8^cre mice. (B) Statistical analysis of brain water content results in adoptive transfer assay (n = 4). (C) Statistical analysis of geotaxis reflex time in adoptive transfer assay (n = 4). (D) Representative H&E staining image (original magnification, ×400) in adoptive transfer assay. (E) Representative Nissl staining image (original magnification, ×400) and (F) statistical analysis of the hippocampal neuronal damage scores in adoptive transfer assay (n = 4). (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 adoptive transfer assay (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 counts of CD68⁺ IBA1⁺ double positive microglia cells within the hippocampal for adoptive transfer assay (n = 4). (K and L) Statistical analysis of CD80/CD206 and CD86/CD206 relative ratios (K), p-STAT1, pNF-κB Thr254 and pNF-κB Ser529 levels of CD11b⁺ CD45^low microglia (L) (n = 5). Data were pooled from two independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001, and ****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, and J-L)