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. 2021 May 13;12:652782. doi: 10.3389/fimmu.2021.652782

Figure 2.

Figure 2

Primary KCs and hepatocytes undergo pyroptosis upon APAP treatment. Primary cells were plated on rat tail collagen-coated plates, and APAP (10 mM/well) was administered after stable adherence. (A) Kupffer cellular supernatant levels of LDH were assessed, n=6. (B) GSDMD-N, caspase-1 p20, IL-1β, and IL-18 protein levels in primary KCs were determined, n=3. (C) Pro-GSDMD/GSDMD-N, pro-caspase-1/caspase-1 p20, pro-IL-1β/IL-1β, and pro-IL-18/IL-18 protein levels in primary hepatocytes were determined, n=3. (D) Hepatocellular supernatant levels of LDH were assessed, n=6. (E) Primary hepatocytes were obtained from untreated C57BL/6 mice and reseeded in a monolayer or the lower chamber of a transwell system. For coculture, primary KCs from untreated C57BL/6 mice were reseeded in the upper chamber of the transwell system. The levels of the indicated proteins (GSDMD-N, caspase-1 p20, IL-1β, IL-18) in the primary hepatocytes from the monolayer or transwell cultures were detected by western blotting, n=3. **P<0.01.