Ld540

HepaRG cell culture and differentiation was performed as described earlier ^{51 (link)}. Differentiated HepaRG (dHepaRG) cells were cultured on 4-well-glass chamber slides (Lab-Tek II, CC2, ThermoFisher Scientific, Bremen, Germany) (5.5x10⁴ cells/well). The cells were stimulated with the fatty acids (opFAs): oleic acid (66 μM) and palmitic acid (33 μM), opFAs opFAs and the interleukin17-alpha (50 ng/ml) (Recombinant Human IL-17-alpha, RnD Systems), or opFAs and [5-(p-Fluorophenyl)-2-ureido]thiophene-3-carboxamide (TPCA1) (5nM) (Sigma Aldrich) in Williams E Medium (William’s Medium E, with stab. glutamine, without Phenol Red, with 2,24 g/l NaHCO3) (PAN Biotech) for 24 h. Cells grown in Williams E Medium without supplement for 24 h were considered as a negative control (CTRL). For each of those conditions, cells were seeded in three different wells which were considered as technical replicates (Table S1). After washing, cells were fixed for 15 min with 4% paraformaldehyde (Sigma Aldrich) at room temperature. Then the cells were washed and stained with Hoechst (1μg/ml) (Hoechst 33342) (ThermoFisher Scientific) and LD540 (0.1 μg/ml) ^{24 (link)} in PBS for 30 min at room temperature. After washing, cells were stored in dH2O at 4 °C for one night maximum.