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. 2013 Aug 15;4(8):e771. doi: 10.1038/cddis.2013.299

Figure 4.

Figure 4

Fas-FasL mediates denitrosylation and activation of procaspase-3 during I/R in hippocampus. (a) Time course analysis of FasL levels in hippocampal CA1 derived from sham-treated rats or rats with 15 min ischemia (I) at various time points (1, 2, 3, 6, and 12 h) after reperfusion (R). n=4; *P<0.05 compared with sham group. (b) FasL AS-ODNs (AS) affects FasL expression in sham rats. 10 nmol FasL AS-ODNs, sense-ODNs control (sense), or vehicle (TE) was administrated to the rats every 24 h for 3 days. FasL-AS significantly decreased FasL expression, whereas sense-ODNs had no effect (n=4; *P<0.05 compared with TE group). (c) FasL AS-ODNs (AS) affects FasL expression and Fas-FasL binding during I/R. 10 nmol FasL AS-ODNs, sense-ODNs control (sense), or vehicle (TE) was administrated to the rats every 24 h for 3 days before ischemia. After 6 h reperfusion, part of the samples were subjected to coimmunoprecipitation analysis of FasL with Fas followed by western blotting with a FasL antibody, the left samples were immunoblotted with FasL, Fas, β-actin antibodies, respectively. FasL-AS decreased both FasL expression and Fas-FasL binding, whereas sense-ODNs had no effect (n=4; *P<0.05 compared with sham group, #P<0.05 compared with TE group). (d) FasL AS inhibited denitrosylation of procaspase-3 after 3 h reperfusion, whereas sense-ODNs had no effect. Samples were processed using the biotin switch method followed by western blotting. n=4; *P<0.05 compared with sham group, #P<0.05 compared with TE group. (e) FasL AS inhibited procaspase-3 activation after 6 h reperfusion, whereas sense-ODNs had no effect (n=4; *P<0.05 compared with sham group, #P<0.05 compared with TE group). (f) FasL AS prevented the upregulation of Trx-2 level induced by I/R. n=4; *P<0.05 compared with sham group, #P<0.05 compared with TE group. Data are represented as means±S.D.