Key Resources Table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-GSDMD-N antibody | Cell Signaling Technology | 36425; RRID: AB_2799099 |
| Anti-GSDMD antibody | Cell Signaling Technology | 96458; RRID: N/A |
| Anti-TMEM173 antibody | Cell Signaling Technology | 13647; AB_2732796 |
| Anti-phospho-TMEM173 antibody | Cell Signaling Technology | 19781; RRID: AB_2737062 |
| Anti-ITPR1 antibody | Abcam | ab5804; RRID: AB_305124 |
| Anti-ACTB antibody | Cell Signaling Technology | 3700; RRID: AB_2242334 |
| Anti-γ-H2AX antibody | Cell Signaling Technology | 9718; RRID: AB_2118009 |
| Anti-cleaved CASP1 antibody | Cell Signaling Technology | 4199; RRID: AB_1903916 |
| Anti-CASP1 antibody | Cell Signaling Technology | 2225; RRID: AB_2243894 |
| Anti-CASP11 antibody | Cell Signaling Technology | 14340; RRID: AB_2728693 |
| Anti-cleaved CASP8 antibody | Cell Signaling Technology | 9496; RRID: AB_561381 |
| Anti-CASP8 antibody | Cell Signaling Technology | 9746; RRID: AB_2275120 |
| Anti-F3 antibody (monoclonal rat IgG2c; clone #355220) | R&D systems | MAB3178; RRID: N/A |
| Anti-rat IgG2c antibody (clone MRG2c-67) | BioLegend | 407602 RRID: AB_345342 |
| HRP-linked anti-mouse IgG secondary antibody | Cell Signaling Technology | 7076; RRID: AB_330924 |
| HRP-linked anti-rabbit IgG secondary antibody | Cell Signaling Technology | 7074; RRID: AB_2099233 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| TUDCA | Sigma-Aldrich | 580549 |
| 2-APB | Sigma-Aldrich | 100065 |
| 4PBA | Santa Cruz Biotechnology | 1821-12-1 |
| Primaxin | Merck | 0006-3514-58 |
| U73122 | Sigma-Aldrich | 662035 |
| BAPTA-AM | Sigma-Aldrich | 196419 |
| Z-IETD-FMK | Selleck Chemicals | S7314 |
| Thapsigargin | Sigma-Aldrich | T9033 |
| Tunicamycin | Sigma-Aldrich | T7765 |
| Cell lysis buffer (10X) | Cell Signaling Technology | 9803 |
| SDS sample buffer (2X) | Thermo Fisher Scientific | LC2676 |
| SuperSignal West Pico Chemiluminescent Substrate | Thermo Fisher Scientific | 34580 |
| SuperSignal West Femto Maximum Sensitivity Substrate | Thermo Fisher Scientific | 34095 |
| RIPA buffer | Cell Signaling Technology | 9806 |
| Phosphatase inhibitor cocktail | Sigma-Aldrich | P0044 |
| Protease inhibitor cocktail | Promega | G6521 |
| Nutrient agar | Becton Dickinson | 213000 |
| Brain heart infusion agar | Becton Dickinson | 211065 |
| Dulbecco’s Modified Eagle’s Medium | Thermo Fisher Scientific | 11995073 |
| RPMI 1640 | Thermo Fisher Scientific | 11875119 |
| Fetal bovine serum | Millipore | TMS-013-B |
| 1% penicillin and streptomycin | Thermo Fisher Scientific | 15070-063 |
| SsoFast EvaGreen Supermix | Bio-Rad | 172-5204 |
| TBST | Cell Signaling Technology | 9997 |
| 5% nonfat dry milk | Cell Signaling Technology | 9999 |
| Protein A agarose beads | Cell Signaling Technology | 9863 |
| Critical Commercial Assays | ||
| Human IFNβ ELISA kit | R&D systems | DIFNB0 |
| Mouse IFNβ ELISA kit | R&D systems | 42400-1 |
| Human F3 ELISA kit | R&D systems | DCF300 |
| Mouse F3 ELISA kit | Abcam | ab214091 |
| Human fibrin ELISA kit | MyBioSource | MBS265263 |
| Mouse fibrin ELISA kit | MyBioSource | MBS706338 |
| Human D-dimer ELISA kit | Abcam | ab196269 |
| Mouse D-dimer ELISA kit | MyBioSource | MBS723281 |
| Human TNF ELISA kit | R&D Systems | DTA00D |
| Human IL1A ELISA kit | R&D Systems | DLA50 |
| Human IL1B ELISA kit | R&D Systems | DLB50 |
| Human IL6 ELISA kit | R&D Systems | D6050 |
| Human HMGB1 ELISA kit | IBL International | ST51011 |
| iScript cDNA Synthesis Kit | Bio-Rad | 1708890 |
| BCA assay kit | Cell Signaling Technology | 7780 |
| Rneasy Plus Kit | QIAGEN | 74134 |
| Fura-2 Calcium Flux Assay Kit | Abcam | ab176766 |
| EasySep Mouse Monocyte Isolation Kit | STEMCELL Technologies | 19861 |
| Experimental Models: Cell Lines | ||
| THP1 | ATCC | TIB-202 |
| HPBM | STEMCELL Technologies | 70042 |
| Tmem173−/− THP1 | InvivoGen | thpd-kostg |
| Tbk1−/− THP1 | InvivoGen | thpd-kotbk |
| Irf3−/− THP1 | InvivoGen | thpd-koirf3 |
| TMEM173-V155M THP1 | InvivoGen | thpd-m155 |
| TMEM173-N154S THP1 | InvivoGen | thpd-s154 |
| Experimental Models: Organisms/Strains | ||
| Tmem173−/− mice | The Jackson Laboratory | 025805 |
| Tmem173flox/flox mice | The Jackson Laboratory | 031670 |
| Lyz2-Cre mice | The Jackson Laboratory | 004781 |
| Lck-Cre mice | The Jackson Laboratory | 003802 |
| GsdmdI105N/I105N mice | Dr. Vishva M. Dixit | N/A |
| cGAS−/− mice | The Jackson Laboratory | 026554 |
| Irf3−/− | RIKEN BioResource Research Center | RBRC00858 |
| Ifnar1−/− | The Jackson Laboratory | 028288 |
| C57BL/6 wild-type mice | The Jackson Laboratory | 000664 |
| E. coli (Serovar O1:K1:H7) | ATCC | 11775 |
| S. pneumoniae (CIP 104225) | ATCC | 6303 |
| Oligonucleotides | ||
| Mouse Zdhhc1 siRNA | Dharmacon | L-056132-01-0005 |
| Mouse Stim1 siRNA | Dharmacon | L-062376-00-0005 |
| Mouse Atp2a1 siRNA | Dharmacon | L-064501-00-0005 |
| Mouse Atp2a2 siRNA | Dharmacon | L-040968-00-0005 |
| Mouse Atp2a3 siRNA | Dharmacon | L-041023-00-0005 |
| Human Zdhhc1 siRNA | Dharmacon | L-014193-01-0005 |
| Human Stim1 siRNA | Dharmacon | L-011785-00-0005 |
| Human Atp2a1 siRNA | Dharmacon | L-006113-00-0005 |
| Human Atp2a2 siRNA | Dharmacon | L-004082-00-0005 |
| Human Atp2a3 siRNA | Dharmacon | L-006114-00-0005 |
| Human Tmem173 siRNA | Dharmacon | L-024333-00-0005 |
| Human Tbk1 siRNA | Dharmacon | L-003788-00-0005 |
| Human Irf3 siRNA | Dharmacon | L-006875-00-0005 |
| See Table S2 for primers used for qPCR | This paper | Table S2 |
| Gsdmd-N cDNA | Dr. Feng Shao | Shi et al., 2015 |
| Gsdmd-C cDNA | Dr. Feng Shao | Shi et al., 2015 |
| Gsdmd-D275A | Dr. Feng Shao | Shi et al., 2015 |
| Atp2a2 cDNA | OriGene | RC207626 |
| Software and Algorithms | ||
| Image Lab software | Bio-Rad | http://www.bio-rad.com/en-us/product/image-lab-software?ID=KRE6P5E8Z |
| CFX Manager software | Bio-Rad | http://www.bio-rad.com/en-us/sku/1845000-cfx-manager-software?ID=1845000 |
| Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |