## 3 Results
### Metabolite Variance across Multiwell Fluidic System to Ascertain Sites of Generated Cell Byproducts
#### 3.8.1 Trending Relevance
(**d3--rat hepatocyte line graph**) NEF stimulates a process shift at upstream well 3, a special cause at wells 7-8, and an unusual outcome in downstream wells 10 and 11. Values a1-d1 represented as the mean \(\pm\) SEM (n = 9). * \(p<0.05\), **\(p<0.01\), *** \(p<0.001\), and *** \(p<0.0001\). Figure 10: LDH release across multiwell cultures on Day 7. LDH activity shown as fold change (vertical axis) vs. DMSO vehicle controls; vehicle normalized at 1. (**a1,b1**) Effect of aspirin exposure in HepaRG cells and rat hepatocytes. (**c1,d1**) Effect of NEF exposure in HepaRG cells and rat hepatocytes. (**a2,b2,c2,d2**) Re-expression of data using individual trending charts (i.e., X control charts) to convey DMSO controls with drug study outcomes. (**a2—HepaRG line graph**) ASA does not induce LDH. (**b2—rat hepatocyte line graph**) ASA stimulates LDH across all wells, 3–11, with increased relevance at upstream well 3 (*) and downstream wells 10 (*) and 11 (*). (**c2—HepaRG line graph**) NEF stimulates LDH at upstream wells 3 (***), 4 (***), 5 (*), and 6 (***). (**d2—rat hepatocyte line graph**) NEF stimulates LDH across all device wells, with increased relevance at upstream well 3 (***), midstream well 7 (***), downstream wells 10 (**) and 11 (*). (**a3,b3,c3,d3**) Variability charts (i.e., MR control charts) to display functional fold-changes amid adjacent wells. (**a3—HepaRG line graph**) ASA does not induce a trending variation. (**b3--rat hepatocyte line graph**) ASA stimulates a process shift at upstream well 3 and special cause in downstream wells 10 and 11. (**c3—HepaRG line graph**) NEF induces a trending process shift at upstream wells 4–7. (**d3—rat hepatocyte line graph**) NEF stimulates a process shift at upstream well 3, a special cause at wells 7–8, and an unusual outcome in downstream wells 10 and 11. Values a1-d1 represented as the mean \(\pm\) SEM (n = 9). * \(p<0.05\), **\(p<0.01\), *** \(p<0.001\), and *** \(p<0.0001\). (wells 6-9). (**b3--rat hepatocyte line graph**) ASA is inconsequential at upstream well 3, stimulates a special cause \"A\" in wells 4-9, then an upregulation special cause (A+) in downstream wells 10 and 11. (**c3--HepARG line graph**) NEF induces a negative trending process shift across the entire \(\upmu\)channel. (**d3--rat hepatocyte line graph**) NEF creates a negative process shift, wells 3-6, normal functions wells 7-8, special cause wells 9-10, and unusual decline outcome in well 11. Values a1-d1 are represented as the mean \(\pm\) SEM (n = 9). * \(p<0.05\), ** \(p<0.01\), and **** \(p<0.0001\). Figure 11: CYP3A4 enzyme activities across multiwell culture locations at Day 7. Activity shown as fold changes (vertical axis) vs. DMSO vehicle control; vehicle normalized at 1. (**a1,b1**) Effect of ASA exposure in HepaRG cells and rat hepatocytes. (**c1,d1**) Effect of NEF exposure in HepaRG cells and rat hepatocytes. (**a2,b2,c2,d2**) Re-expression of data using individual trending charts (i.e., X control charts) to convey DMSO controls with drug study outcomes. (**a2—HepaRG line graph**) ASA impedes CYP3A4 at upstream wells 3 (**) then shows upregulated stimulation in midstream wells 6 (*), 7 (*), 8 (*), and 9 (*). (**b2—rat hepatocyte line graph**) ASA has no effect on upstream well 3, then stimulates CYP3A4 across all remaining downstream wells 4 (*)–9 (*) and 10 (****)–11 (****). (**c2—HepaRG line graph**) NEF inhibits CYP3A4 activity across the entire \(\upmu\)fluidic channel, inclusive of wells 3–11. (**d2—rat hepatocyte line graph**) NEF inhibits CYP3A4 activity in upstream wells 3–6 (*), regains normal function in wells 7–8, has functional increases in wells 9 (*) and 10 (****), and returns to near normal activity in downstream well 11. (**a3,b3,c3,d3**) Variability charts (i.e., MR control charts) to display functional fold changes amid adjacent-wells. (**a3—HepaRG line graph**) ASA induces a trending process shift of upstream inhibition (well 3) followed by midstream recovery (wells 6–9).