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Additional, more stringent wash buffers are used to remove hydrophobically bound and precipitated proteins

An inverted fluorescence microscope (Nikon, DS-Ri2, Japan) was used to obtain fluorescence images

Heavy metals: Are arsenic, cadmium, lead and mercury within appropriate limits

1 Introduction The unprecedented pace of industrialization and urbanization over recent decades has introduced a vast array of chemical contaminants into our environment, among which environmental endocrine disruptors (EEDs) have emerged as a particularly concerning threat to human reproductive health ( 1.1 Sources and classification of EEDs Environmental endocrine disruptors encompass a remarkably diverse array of chemical compounds that can be systematically categorized into three distinct groups based on their origin, chemical properties, and environmental behavior: Heavy Metals and Metalloids represent one of the most persistent categories of reproductive toxicants