DMSO-reconstituted methanolic extracts were shipped to Attagene, Inc. (Morrisville, NC, USA) for testing by their TF-FACTORIAL (45 TF specific reporters) and NR-FACTORIAL (24 human NRs) assays (previously named cis-and trans-FACTORIAL assays, respectively). The assays use HepG2 cells to assess the activity of endogenous transcription factors (TF) or transfected hybrid proteins consisting of a yeast GAL4 DNA binding domain and ligand-binding domain of the human nuclear receptors (NR). These multiplexed assays comprised 69 measured endpoints (Table ) related to cell stress, endocrine activity, growth and differentiation, immunity, and lipid, xenobiotic, and general metabolism. Extracts were tested at a single concentration (3 µL DMSO extract/mL cell culture medium) for 24 h for the NR-FACTORIAL assay at three concentrations (1, 3, and 9 µL DMSO extract/mL cell culture medium) for 24 h for the TF-FACTORIAL assay. The TF-FACTORIAL assay was repeated twice at the midpoint concentration. Final DMSO concentrations were 0.1-0.9% (v/v), depending on the concentration of extract used in the assay. Three to six technical replicates of DMSO solvent controls matched to the DMSO concentration of the extracts were run with each sample set. Each extract was run as three technical replicates in Dulbecco's Modified Eagle Medium (DMEM) containing 1% charcoal-stripped fetal bovine serum (FBS). Reporter RNA was isolated, amplified by reversetranscription polymerase chain reaction (RT-PCR), labeled with fluorescent markers, and quantitively assayed by capillary electrophoresis. Bioassay responses were expressed as foldinduction relative to the DMSO control by dividing the treated cells' average technical replicate expression by the average technical replicate expression of the appropriate DMSO control.