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After being blocked for 60 minutes with blocking buffer (5% normal donkey serum, 1% BSA, 0.3% Triton X-100), sections were incubated in primary antibodies CD31 for immunofluorescence (1:100, BD Biosciences, 558736) and p-eNOS (1:100, Cell Signaling Technology, 9571s) overnight at 4°C

then tap lightly for absorption