Figure 5. Methane reduced apoptosis induced by sepsis. The lung and intestine tissues were harvested 12 and 24h after cecal ligation and puncture (CLP) (n = 12). (A) Representative TUNEL staining in lung and intestine tissues (Scale bars: 50μm). (B) The percentage of TUNEL positive cells were measured. (C) Representative Western blot and quantitative analysis of (D) Cytochrome c, (E) caspase-3, (F) caspse-9 in lung tissue, RAW264.7 cells and primary peritoneal macrophages. (G-H) Flow cytometry analysis of RAW264.7 cell apoptosis percentage at 24 h after the stimulation of lipopolysaccharide+ adenosine-triphosphate (LPS+ATP) (n = 3). (Data are shown as the mean ±SD. ^*P < 0.05; ^**p < 0.01; ^***P < 0.001; ^#P < 0.05 versus normal medium (NM) group; ^$p < 0.05 versus methane-rich medium (MRM) group; ^&P < 0.05 versus. normal medium+ lipopolysaccharide+ adenosine-triphosphate (NM+L+A) group).