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. 2020 Mar;8(5):168. doi: 10.21037/atm.2020.02.21

Figure 3.

Figure 3

NF-kB-NLRP3 signaling involves in PA-induced HSCs phenotype change. Serum-starved HSCs (day 2) were transfected with lentivirus vector containing NLRP3 shRNA (LV452_pL-NLRP3-shRNA-3300) for 48 h or were pre-incubated with NF-kB inhibitor CAPE (1 µm/mL) for 2 h then exposed to PA (200 µm) combined with LPS (1 µg/mL) for 12 h. (A,B) Gene and protein NLRP3 relative expression level was evaluated by PCR and densitometric analysis, respectively; (C) cells were observed under fluorescence and optical microscope. (D) Gene expression of IL-1β, Caspase 1 and IL-18 were determined by qPCR. (E) IL-1β and IL-18 level in cell supernatant were detected by ELISA; HSCs phenotype was evaluated by assessing fibrotic markers (F), HSCs proliferation (G), migration (H) and apoptosis (I) as described in Methods. Results are presented as mean ± standard error. *, P<0.05, **, P<0.01 compared with control (untreated). (C) Magnification ×200; (H) 100 µm.