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Membranes were washed with 0.1% Tween-20 and 1 PBS 3 times and incubated with horseradish-peroxidase-conjugated secondary antibodies (Jackson Laboratories) in 5% non-fat skimmed milk powder (Rgilait) in 0.1% Tween-20 and 1 PBS for 1 h at room temperature and washed 3 times with 0.1% Tween-20 and 1 PBS

The rationale is that effects accumulate during the course and persist between courses, with new courses reinforcing the previous effects

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It is safe when it is advised by your healthcare professionals