Fig. 3.
Characterization of immortalized Kupffer cells (iKC). iKC from three different passages were seeded and cultured for 48 h with 2% DMSO. KC purified from three different donors were seeded and cultured for 24 h. THP1 were seeded and cultured for 48 h with PMA. Cells were then left untreated (a, c), exposed to pHrodoTM bacteria for 1 h at 4°C or 37°C (b) or exposed to the indicated ligands at the concentration indicated in online supplementary Table S3 for 24 h (d). a, bi Microscopic analyses of cells (×20 magnification) were performed. bii Bacteria phagocytosis was assessed by flow cytometry analysis. c Proteins were extracted, pooled and TLR1 to TLR9, MyD88, TRIF, RIG-I, MDA-5, MAVS, cGAS, IFI16, STING, and PD-L1 protein expression was assessed by Western blot analyses. Target protein levels are normalized to total protein quantification assessed by stain-free staining. Stimulated cells were used as controls (CTR+) for primary antibody efficiencies as indicated in online supplementary Table S2. d Supernatants were collected, and IL-6 and IP-10 secretions were analyzed by ELISA. Data are presented as mean ± standard deviation of three biological replicates.