Supplementary Materials

ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation

Sebastian Rühl, Kateryna Shkarina, Benjamin Demarco, Rosalie Heilig, José Carlos Santos, Petr Broz

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S18
  • Captions for Movies S1 to S8

Images, Video, and Other Media

Movie S1
Pam3CSK4-primed wildtype (left) and Casp11-deficient (right) BMDMs were loaded with 4 μM Fluo-8 (green) and subsequently transfected with LPS. Images were acquired in the presence of PI (red) every 30 seconds. Time is shown in minutes:seconds.
Movie S2
Pam3CSK4-primed wildtype (left) and Gsdmd-deficient (right) BMDMs were loaded with 4 μM Fluo-8 (green) and subsequently transfected with LPS. Images were acquired in the presence of PI (red) every 30 seconds. Time is shown in min:seconds
Movie S3
CHMP4-GFP HeLa cells stably transduced with Dox-inducible GSDMDNT were treated with doxycycline for 3 hours and subsequently imaged. Images were acquired every 10 seconds in the presence of PI (red). Time is shown in minutes:seconds.
Movie S4
Related to Figure 2B. Time-lapsed confocal images of either CHMP4-mCherry-expressing DmrB-Casp-1- transgenic HEK293T cells stained with Annexin-V as membrane marker. A cell expressing high level CHMP4 was chosen for imaging. B/B homodimerizer was added at t=0.
Movie S5
Related to Figure 2C. Time-lapsed confocal images of either CHMP4-GFP (in red)-expressing DmrB-Casp- 1-transgenic HEK293T cells stained with Annexin-V as membrane marker. A cell expressing high level CHMP4 was chosen for imaging. B/B homodimerizer was added at t=0.
Movie S6
Related to Figure S5A. Time-lapsed confocal images of either CHMP4-mCherry-expressing DmrB-Casp- 1-transgenic HEK293T cells stained with Annexin-V as membrane marker. A cell expressing high level CHMP4 was chosen for imaging. B/B homodimerizer was added at t=0.
Movie S7
Related to Figure S9. VPS4AWT (GFP-labeled)- and VPS4AE228Q (unlabeled) expressing immortalized WT macrophages were transfected with LPS and imaged in the presence of PI (red) every 5 minutes.
Movie S8
Related to Figure S9. VPS4AWT (unlabeled)- and VPS4AE228Q (GFP-labeled) expressing immortalized WT macrophages were transfected with LPS and imaged in the presence of PI (red) every 5 minutes.