Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2021 Jun 2.

Published in final edited form as: J Pharmacol Toxicol Methods. 2018 Mar 16;92:77–94. doi: 10.1016/j.vascn.2018.03.002

Figure 7. — HEK293T cells were transfected with 25 ng of βgalactosidase or 24 ng of CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, or a 10-CYP liver mix (Table 1 for ratio) + 1 ng P450 oxidoreductase mRNA at a lipid:mRNA ratio of 2.5 and delivered at 5 μL per well. Fifty-six test compounds (Table S5) were delivered at a volume of 225 nL with an acoustic dispenser (0.5% DMSO final) using randomized plate designs at final concentrations ranging from 1–100 μM (x-axis) at 6 hours post-transfection and incubated for 36 hours. Cytotoxicity (y-axis) was measured in three replicate experiments and normalized data fit with a 3-parameter Hill model (bottom constrained to zero, dashed black line). Red dashed line showed the mean ± 2*sd for the DMSO vehicle control wells. Shown are example curves for aflatoxin B1 (A), amodiaquin (B), amitripytyline (C), 8-quinolinol (D), azobenzene (E), and fenazaquin (F). All ten CYP singlets and liver mix were run for each chemical; however, for clarity only βgal, liver mix and those CYP singlets showing bioactivation or detoxification were plotted. See Supplemental Figure 1 for all curves and bio-groups.

Figure 7. — HEK293T cells were transfected with 25 ng of βgalactosidase or 24 ng of CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, or a 10-CYP liver mix (Table 1 for ratio) + 1 ng P450 oxidoreductase mRNA at a lipid:mRNA ratio of 2.5 and delivered at 5 μL per well. Fifty-six test compounds (Table S5) were delivered at a volume of 225 nL with an acoustic dispenser (0.5% DMSO final) using randomized plate designs at final concentrations ranging from 1–100 μM (x-axis) at 6 hours post-transfection and incubated for 36 hours. Cytotoxicity (y-axis) was measured in three replicate experiments and normalized data fit with a 3-parameter Hill model (bottom constrained to zero, dashed black line). Red dashed line showed the mean ± 2*sd for the DMSO vehicle control wells. Shown are example curves for aflatoxin B1 (A), amodiaquin (B), amitripytyline (C), 8-quinolinol (D), azobenzene (E), and fenazaquin (F). All ten CYP singlets and liver mix were run for each chemical; however, for clarity only βgal, liver mix and those CYP singlets showing bioactivation or detoxification were plotted. See Supplemental Figure 1 for all curves and bio-groups.