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. 2016 May 12;38(1):161–171. doi: 10.3892/ijmm.2016.2586

Figure 6.

Figure 6

Evaluation of the expression of apoptosis signal-regulating kinase 1 (ASK1), c-Jun N-terminal kinase (JNK), phosphorylated (p-)JNK, pro-caspase-3 and cleaved caspase-3 by western blot analysis and RT-qPCR. Representative blots are shown. β-actin or GAPDH were used as controls. Levels of particular transcripts were quantified by real-time PCR using gene-specific primers. The amount of each target transcript was normalized by measuring β-actin transcript levels. (A) Pro-caspase-3 levels in the hypothermic machine perfusion (HMP) group were significantly reduced compared with those in the cold storage (CS) group (*P<0.05) and the cleaved caspase-3 levels in the HMP group were significantly lower than those in the CS group (**P<0.01). Compared with the sham group, both the HMP and CS groups showed increased expression of cleaved caspase-3 (both **P<0.01). (B and D) The HMP group exhibited reduced ASK1 expression at both the protein and mRNA levels compared with the CS group (**P<0.01). (C) Compared with the sham group, both the HMP and CS groups showed reduced expression of total JNK, although this effect was not statistically significant. The expression of p-JNK was significantly reduced in the HMP group compared with that in the CS group (**P<0.01). (E) Compared with the sham and CS groups, the mRNA expression of A20 was significantly increased in the HMP group (**P<0.01). Data represent the means ± SD of three experiments.