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The membrane was blocked with 5% delipidated milk in PBS containing 0.1% Tween 20 (PBST) for 1 h, incubated with polyclonal anti-PEDF antibodies (#PAB972Hu01, Cloud-Clone corp., China, 1:1000 in PBST) for 1 h, washed with PBST, and incubated with horseradish peroxidase-conjugated anti-rabbit IgG antibody (#111-035-003, Jackson ImmunoResearch, UK, 1:50,000 in PBST) for 1 h