Figure 4

Endpoint-specific Sensitivity to 6-Aminonicotinamide. (a-f) Beas-2B cells treated with graded concentrations of 6AN and B[a]P for 48 h were assessed for (a) extracellular leakage via LDH and for (b) ΔΨm via JC-1, (c) Alamar Blue, (d) MTT, (e) MTS, and (f) WST1 metabolism. Vehicle- and inhibitor-only controls are represented by a unified group designated as R. Ctrl. n = 3. (g,h) Cells treated with graded concentrations of 6AN were assessed for cytotoxicity via (f) extracellular LDH leakage and (g) resazurin/tetrazolium endpoint metabolism as well as reduction in cell number as indicated by total protein recovery post-treatment; curve designations are annotated within the graph. n = 3. (i) Normalized endpoint metabolism induced by 10 µM B[a]P regressed against baseline endpoint-specific metabolic reduction by 0.5 µM 6AN demonstrated excellent linear correlation (Coefficient of correlation 95.4%). (j) Beas-2B cells treated with 6AN were assessed for NADPH content with correction for total protein. n = 3. (k–m) Beas-2B cells treated with 1 µM or 10 µM B[a]P for 48 h in the presence of 0.5 µM 6AN, followed by (k) EdU uptake in EdU + cells, (l) cumulative cell cycle phase quantitation, and (m) magnitude changes in each cell cycle relative to respective control. n = 3. Endpoints are the arithmetic mean of listed independent experiments with error bars denoting one standard error of the mean.