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After 30 min, the lysate was clarified by centrifugation (1500 g ), filtered (0.22 m filter), aliquoted and the E1E2-containing lysates were stored at 80 C

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2.15 Real-time quantitative PCR Total RNA was extracted from aortic tissue and endothelial cells using TRIzol reagent (Takara Bio Inc., 9108), following the manufacturers instructions

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