Figure 4
GSDMD-N but not GSDMD-C forms oligomer during pyroptosis. (A) Gsdmd−/− RAW-asc cells were reconstituted with Flag-GSDMD and GSDMD-Flag respectively, and then primed with LPS for 4 h followed by Nig stimulation for 0 min, 30 min, 60 min and 120 min. The culture supernatants together with their corresponding cell extracts were resolved by 4% - 12% gradient SDS-PAGE. Immunoblotting of Flag was performed. (B) The same cells were treated as in A. The culture supernatants together with their corresponding cell extracts were analyzed after DSS crosslinking. (C) Flag-GSDMD-reconstituted cells were primed with LPS followed by Nig stimulation or no stimulation. Lysates of these cells were fractionated by gel-filtration chromatography (Superose 6, fractions sizes at top) and followed by immunoblot analysis.
