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The specific procedures for the respective assays are described in detail below.
The minimum detection limit for the respective assays was PlGF: 3.9 pg/ml, sEng: 62.5 pg/ml, sFlt-1 31.3 pg/ml.
Wild-type strains not harboring a plasmid were grown in the presence of 0.07% ethanol in the respective assays, when compared to the strains harboring a plasmid to rule out an effect of the ethanol.
This solvent concentration had no effect in the respective assays.
*All positive samples confirmed in their respective assays.
These values represent the lower VL values at which the respective assays keep their linearity.
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Interestingly, the activities of NADH oxidase were dependent on availability of oxygen in the respective assay reactions.
In each sub-Figure, the left column shows a representative image of the respective assay, and the right column shows the quantification of the assay.
In an attempt to standardise the effects of the different estrogens in receptor-based assays, their relative estrogenic potency (REP) has been defined in relation to the effect of E2 in the respective assay (e.g. [23]).
The respective assay was capable of detecting 0.125 pg/mg stanozolol and 0.25 pg/mg 3′-hydroxystanozolol with 50 mg hair; 0.063 ng/mL stanozolol and 0.125 ng/mL 3′-hydroxystanozolol with 100 μL of urine or serum.
The assays show optimal performance characteristics in their validation on plasmids containing the respective assay target sequences.
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