Fig. 9
LPS induces microglial programmed cell death in neonatal mice at 24 h. A. Heatmap displaying q-values from the KEGG cellular process enrichment analysis with significant pathways indicated (*P < 0.05, ***P < 0.001, NS vs. LPS). B. Heatmap showing differential gene expression related to the programmed cell death signaling pathway in neonatal mice exposed to 10 mg/kg LPS. C. PPI network of of cell death regulatiory proteins, extracted from the KEGG and STRING databases. Line thickness represents interaction confidence levels (thicker lines indicate higher confidence). D. RT-PCR validation of key programmed cell death genes in mouse brain tissue 24 h after LPS injection. Statistical tests used include one-way ANOVA with Tukey’s post hoc test or Kruskal–Wallis test with post hoc Dunn test (Mlkl: H = 10.4, P = 0.002; Zbp1: H = 5.4, P = 0.02; Ripk3: F2,13 = 9.5, P = 0.002; Ripk2: H = 8.6, P = 0.013; Slc7a11: F2,13 = 5.0, P = 0.02; Slc40a1: F2,13 = 17.0, P = 0.000. n = 4–8/group). Significant results: &P < 0.05, NS vs. 5 mg/kg LPS, * P < 0.05, ** P < 0.01, *** P < 0.001, NS vs. 10 mg/kg LPS, ##P < 0.01, 5 mg/kg LPS vs. 10 mg/kg LPS. E. Representative electron microscope images of mitochondria in control and LPS-treated mice at 24 h after 10 mg/kg LPS treatment. Red and blue arrows indicate normal and damaged mitochondria, respectively (Scale bar = 200 nm). n = 3–4/group. F. Representative TUNEL (green) and Iba-1 (red) double-staining images of the 10 mg/kg LPS group at 24 h. Scale bars, 50 μm (upper panel) and 20 μm (lower panel)