Figure 2.

DPYD inhibition reduces lipid burden in an in vitro model of steatosis. (A) The inhibition of DPYD led to increased uracil levels in media collected from treated PHHs with an IC50 of 56 nM. (B) While an increase in LipidTox signal intensity is observed in the presence of 200 µM FFAs, this effect is diminished in the presence of 250 nM Gimeracil, a DPYD inhibitor. Scale bar represents 100 µM. (C) Quantification of lipid droplet accumulation in the hepatocytes with an Operetta Harmony image analysis pipeline (spot area per cell area) highlights a more than twofold increase in lipid droplet area with exposure to FFAs (square) relative to vehicle treatment (circle). Gimeracil exposure significantly reduces lipid droplet accumulation in both the FFA and vehicle control conditions in a dose responsive fashion. The degree to which Gimeracil inhibits lipid accumulation in the presence of fatty acids is represented by an IC50 value of 247.8 nM with an R² of 0.8481. (D) Cell viability is not significantly impacted by either FFA exposure (200 µM) or DPYD inhibitor up to a concentration of 500 nM as measured by a calcein AM stain confluence quantified by the Incucyte Zoom analysis software. (E) Neither 200 µM FFA exposure or DPYD inhibition in the range of 50–500 nM is cytotoxic as measured by Ethidium homodimer-1 staining and quantification of confluence with the Incucyte Zoom analysis software.