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. 2017 Mar 28;8(25):40741–40755. doi: 10.18632/oncotarget.16580

Figure 11. Effect of antioxidant NAC and cathepsin B inhibitor CA-074 Me in mitochondrial apoptosis pathway in vivo.

Figure 11

BALB/c mice were pretreated with or without100mg/kg N-acetyl-L-cysteine (NAC, intraperitoneal) at 1h or 10mg/kg 50μM CA-074 Me (intraperitoneal) at 30min followed by exposure in a temperature controlled chamber (ambient temperature 35.5±0.5°C and 60±5% relative humidity)until the rectal core temperature (Tc)reached 42°C. A. TUNEL staining of the ileum of mice (magnification, ×200). B. Quantification of TUNEL-positive index. C. Cytochrome C was determined by Western blots. β-actin was run as an internal control. COX IV was used as a mitochondrial loading control. D. Quantification of Western blots for cytochrome C. E. Bcl-2 and Bax were determined by Western blots. β-actin was run as an internal control. F. Quantification of Western blots for Bax/Bcl-2 ratio. G. Enzymatic activity of caspase-9 was measured in cell lysates using a fluorogenic substrate, Ac-LEHD-AFC. H. Enzymatic activity of caspase-3 was measured in cell lysates using a fluorogenic substrate, Ac-DEVD-AMC. *P < 0.05, statistically significant relative to control (37°C),#P < 0.05, statistically significant relative to heat stress group (HS),(n=6/group).