Fig. 1
Pro-fibrotic markers and NLRP3 were both reduced by MCC950 in LX-2 cells triggered by SEA. a Morphology of LX-2 cells in different treatment groups observed under a microscope. b The mRNA and protein expressions of NLRP3, α-SMA and Collagen I were determined by RT-PCR and western blot, respectively. Relative quantitative western blot analysis of NLRP3, α-SMA and Collagen I was performed. Each immunoreactive band was digitized and normalized to loading control. c LX-2 cells were incubated with LPS (50 and 100 ng/ml) (top) or SEA (10, 20, 40, 80 μg/ml) (middle) for indicated time. LX-2 cells were pre-incubated with MCC950 (0.1, 1, 10, 100 μM) (bottom) for 0, 1, 4, 8 h, and then subjected to LPS (100 ng/ml) treatment for another 2 h. NLRP3 mRNA expression was determined by RT-PCR. *, #, aP < 0.05, **, ##, aaP < 0.01, ***, ###, &&&, aaaP < 0.001. d MCC950 (1 μM) pre-treatment for 4 h. This was then supplemented with SEA in LX-2 cells for an additional 24 h. LX-2 cells were treated with LPS for 2 h. Immunofluorescence staining of NLRP3 and α-SMA. Immunofluorescence staining of NLRP3 was digitized and normalized to control. Three independent experiments were performed, and one representative figure is shown (b, d). *, #P < 0.05, **P < 0.01, ***, ###P < 0.001 vs PBS; bP < 0.01, bbP < 0.01 vs SEA. Abbreviation: NS, no statistical significance. Scale-bars: a, 200 μm; d, 100 μm