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Embryonic zebrafish were examined for changes in protein expression following exposure to sublethal concentrations of 17β-estradiol (E2) and the estrogen mimic 4-nonylphenol (4-NP). Protein Expression Signatures were derived from embryo homogenates by two-dimensional electrophoresis and digital imaging. In both experiments approximately 30% of the proteins sampled were specific to either E2 or 4-NP and about 33% were common to the control, 4-NP and E2. However, of the proteins induced by either E2 or 4-NP, 28% were common to both chemicals at 1 ppm but only 7% were common to both at 0.1 ppm. While there are many proteins that respond specifically to each chemical, relatively few are common to the two chemicals suggesting that the response pathways of the two chemicals are distinct.