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Analysis of HHEX expression by immunohistochemistry in WT male and female adrenals revealed a striking lower staining intensity in zF cells of female adrenals (Fig

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They were permeabilized with 0.1% Triton X-100 for 15 min and then incubated with 25 ng/mL RNase for 1 h at 37 C to remove unwanted RNA contamination and enhance the specificity of the staining