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Whole blood was collected from animals [males displaying macroscopic adenomas (A, $n=7$ ), VEH controls (V, $n=7$ ), and TBT-treated males without any macroscopic adenomas (T, $n=5$ )] at the time of euthanasia (45 weeks of age) via cardiac puncture, and plasma was isolated by centrifugation at 4°C for 10 min at $500\times g$ after red blood cell lysis with EL Buffer (Qiagen, Catalog No. 79217) and stored at $-80°\mathrm{C}$ until analysis. Alanine and aspartate transaminase (ALT/AST) activity levels were analyzed in the plasma of 45-week-old animals using an alanine transaminase activity assay (ab105134; Abcam) or an aspartate transaminase activity assay (MAK055; Sigma) as per the manufacturer’s instructions. Briefly, plasma was diluted (1:5 for ALT and 1:10 for AST) and mixed with Reaction Mix, then read on the BioTek EL808 plater reader (BioTek, Instruments) in a kinetic fashion every 3 min for 60 min while incubating at 37°C. For analysis two time points (T) were chosen where all samples fell within the standard curve. Pyruvate/glutamate levels were calculated from a standard curve and activity (in nanomoles per minute per milliliter or in milliunits per milliliter) was calculated as $\left(\mathrm{B}/\text{DT}\times \mathrm{V}\right)\times \mathrm{D}$ for ALT or $\left(\mathrm{B}\times \mathrm{D}\right)/\left(\text{DT}\times \mathrm{V}\right)$ for AST, where $\mathrm{B}=\text{pyruvate}/\text{glutamate \hspace{0.17em}}\left(\text{nmol}\right)$ , $\mathrm{\Delta }\mathrm{T}=\mathrm{T}2-\mathrm{T}1$ , $\mathrm{V}=\text{volume\hspace{0.17em}of\hspace{0.17em}original\hspace{0.17em}sample}$ , and $\mathrm{D}=\text{dilution\hspace{0.17em}factor}$ . From this data, the ALT/AST ratio was calculated.