Figure 4.

MiR-34a affects stroke outcomes via interacting with cytochrome c. (A) WES system image showing CYC, VDAC and GAPDH expression from hemispheres of WT and miR-34a^−/− mice at 6 h post-stroke. (B) Relative CYC expression normalized to GAPDH from the data generated by the WES system. CYC level is significantly decreased in ischemic hemispheres of WT mice but no significant changes in miR-34a^−/− mice between contralateral hemispheres (Control) and ischemic hemispheres (Ischemia). N = 5 per group, **p < 0.01, One-way ANOVA followed by post hoc Tuckey’s test was used for data analysis. Data are expressed as mean ± S.D. (C) Relative VDAC expression normalized to GAPDH from the data generated by the WES system. VDAC was not significantly altered in WT mice nor miR-34a^−/− mice. (D) Multiplexed WES system image showing CYC and GAPDH expression from purified pCECs of WT and miR-34a^−/− mice (n = 10 per group, pooled cell samples) at 6 h post-stroke. (E) Relative CYC expression by normalization to GAPDH. A 2.8 fold decrease of CYC level was observed in WT mice but no changes were observed in miR-34a^−/− mice between contralateral hemispheres and ischemic hemispheres. (F) A CYC reporter was coexpressed with a miR-34a plasmid, a miR-34a mimic, a miR-34c mimic, or a plasmid control in cultured cerebral vascular endothelial cells for 24 hours. Relative firefly luciferase activity was evaluated and normalized to renilla luciferase activity. Relative firefly luciferase activity was reduced by miR-34a plasmid and miR-34a mimic. The experiment was repeated 3 times and triplicates were used for each analysis. Data represents the mean ± S.D. *p < 0.05. One-way ANOVA followed by post-hoc Tuckey’s test was used for analysis.