Fig. 4: The supernatants from LPS/NIG-treated caspase-1/11−/− or Gsdmd−/− macrophages cannot cause pyroptosis of ASCs. | Cell Death & Disease

Fig. 4: The supernatants from LPS/NIG-treated caspase-1/11−/− or Gsdmd−/− macrophages cannot cause pyroptosis of ASCs.

From: Induction of ASC pyroptosis requires gasdermin D or caspase-1/11-dependent mediators and IFNβ from pyroptotic macrophages

Fig. 4

a, b WT BMDMs, caspase-1/11−/− BMDMs, WT iBMDMs, and Gsdmd−/− iBMDMs were stimulated with 1 μg/ml LPS for 5 h prior to challenging with 10 μM nigericin for 1 h. The supernatants were collected for detecting caspase-1 p20 by immunoblotting analysis (a, b). Data are representative of at least three independent experiments. c–h Treat LPS-primed ASCs with the above supernatants for 2 h. After the treatments of related supernatants, collect ASCs for immunoblot analysis (out of at least three performed) of GSDMD and IL1β (c, f) or replace the supernatant with fresh media for another 2 h. Harvest the medium for evaluating LDH (d, g) and mature IL-1β release levels (e, h) of ASCs. n = 3. Statistical significance between the control group and treated groups was determined using one-way analysis of variance. Data are shown as mean ± SD. ns: p > 0.05, ***p < 0.001, ****p < 0.0001. LPS lipopolysaccharide, NIG nigericin, ASCs adipose tissue-derived mesenchymal stem cells, BMDMs Bone marrow-derived macrophages, iBMDMs immortalized bone marrow-derived macrophages, WT wildtype, GSDMD gasdermin D, GSDMD-NT N-terminal gasdermin D, SN supernatant, SCB supernatant collected from BMDMs, SLNB supernatant collected from LPS/NIG-treated BMDMs, LDH lactate dehydrogenase.

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