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For IF staining, fixed with 4% PFA for 15 min, permeabilized with 0.3% Triton X-100 in PBS for 10 min, and blocked with 5% normal donkey serum for 1 h at room temperature (RT)

Some cosmetic pigments and lighter inks may respond unpredictably and can darken in certain cases due to oxidation

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Stocco D, X W, Y J, Pr M (2005) Multiple signaling pathways regulating steroidogenesis and steroidogenic acute regulatory protein expression: more complicated than we thought