List of reagents used in the study
| . | . | . |
|---|---|---|
| Antibody | Catalog reference | Provider |
| Rabbit anti-ZAK antibody 1:1,000 | A301-993A | Bethyl Laboratories |
| P38 MAPK antibody 1:1,000 | 9212S | Cell Signaling |
| Phospho-P38 MAPK (Thr180/Tyr182) (D3F9) XPRabbit mAb 1:1,000 | 4511S | Cell Signaling |
| JNK1 Rabbit anti-human polyclonal 1:1,000 | 44690 G | Invitrogen |
| Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb 1:1,000 | 4668S | Cell Signaling |
| Anti-NLRP1 1:200 | 679802 | Biolegend |
| Anti-NLRP1 (N-terminal) 1:1,000 | AF6788-SP;RRID: AB_2916167 | R&D Systems |
| Anti-NLRP1 (C-terminal), 1:1,000 | ab36852;RRID: AB_776633 | Abcam |
| Anti-Sheep igG HRP (1/4,000) | HAF016;RRID: AB_562591 | R&D |
| Goat-anti-rabbit IgG (H+L), HRP conjugate (1/5,000) | R-05072-500;RRID: AB_10719218 | Advansta |
| GSDMDC1 antibody 1:1,000 | NBP2-33422 | Biotechne |
| Anti-puromycin antibody, clone 12D10 | MABE343 | Sigma-Aldrich |
| EEF2 antibody (1:1,000) | 2332S | Cell Signaling |
| DPH1 antibody (1:1,000) | #H00001801-M02 | Thermo Fisher Scientific |
| Anti-α Tubulin antibody 1:1,000 | ab4074 | Abcam |
| Streptavidin—HRP | 434323 | Thermo Fisher Scientific |
| Anti-NLRP3/NALP3,mAb (Cryo-2) | AG-20B-0014-C100 | AdipoGen |
| Reagents | Catalog reference | Provider |
| Transwell | 140620 | Thermo Fisher Scientific |
| NAD+, biotin-labeled | 80610 | BPSBioscience |
| Pseudomonas exotoxin A 0.5 mg | P0184-.5 MG | Sigma |
| Inactive exotoxin A | N.A. | Creative Diagnostics |
| Diphtheria toxin | D0564-1MG | Sigma-Aldrich |
| Cholix toxin | This study | This study |
| Anisomycin | SE-S7409-10 MG | Selleck |
| Talabostat (Val-boro-pro, VbP) | tlrl-VbP-10 | Invivogen |
| PhoSTOP | 4906845001 | Sigma-Aldrich/Roche |
| cOmplete, Mini, EDTA-free protease inhibitor cocktail | 4693159001 | Sigma-Aldrich/Roche |
| Lipofectamine LTX | 15338030 | Invitrogen |
| Puromycin dihydrochloride | A1113803 | Gibco |
| Molecular probes SYTOX Green nucleic acid stain | S7020 | Thermo Fisher Scientific |
| Propidium iodide | P1304MP | Thermo Fisher Scientific |
| Nate | lyec-nate | Invivogen |
| Phos-tag Acrylamide | AAL-107 | Wako Chemicals |
| Manganese chloride(II) | 63535 | Sigma-Aldrich |
| Prestained protein size marker III | 230-02461 | Wako Chemicals |
| Ivacaftor (VX-770) 5 mg/10 µM | S1144 | Selleck |
| Tezacaftor (VX-661) 5 mg/10 µM | S7059 | Selleck |
| Elexacaftor (VX-445) 5 mg/5 µM | S8851 | Selleck |
| Critical commercial assays | Catalog reference | Provider |
| Human total IL-18 duo set | DY318-05 | R&D |
| Cyquant LDH | C20301 | Thermo Fisher Scientific |
| Human IL-1β | 88-7261-77 | Thermo Fisher Scientific |
| Q5 site-directed mutagenesis kit protocol | E0554 | NEB |
| Chemicals | Catalog reference | Provider |
| ZAK inhibitor (PLX-4720) 10 µM | HY-51424 | Med Chem Express |
| P38 inhibitor SB 203580 10 µM | HY-10256 | Med Chem Express |
| Caspase-1 inhibitor—Ac-YVAD-cmk 20 µM | inh-yvad | Invivogen |
| Z-VAD-FMK 20 µM | vad-tlrl: | Invivogen |
| Caspase-3/7 inhibitor ZDEVD-FMK 20 µM | S7312 | Selleck |
| Caspase-8 inhibitor - Z-IETD-FMK | inh-ietd | Invivogen |
| Bortezomib 1 µM | S1013 | Selleck |
| CHX | C4859-1Ml | Sigma-Aldrich |
| MCC950, 10 mg | Inh-mcc | Invivogen |
| hPNECs | Catalog reference | Provider |
| Tryple express enzyme (1X), no phenol red | 10718463 | Thermo Fisher Scientific |
| Nutragen type I collagen solution, 6 mg/ml (Bovine) | 5010-50 Ml | Advanced Biomatrix |
| Sputolysin | 560000-1SET | Sigma-Aldrich |
| PneumaCult-Ex plus medium | 05040 | Stem Cell |
| Costar 6.5 mm transwell, 0.4 µm pore polyester membrane inserts | 38024 | Stem Cell |
| hPCECs | Catalog reference | Provider |
| Human corneal epithelial cell growth medium | 221-500 | Tebu-bio |
| Human corneal epithelial cells | 630-05a | Tebu-bio |
| Cell lines | Catalog reference | Provider |
| A549 ASC GFP NLRP1 | a549-ascg-nlrp1 | Invivogen |
| A549 ASC GFP | a549-ascg | Invivogen |
| A549 ASC GFP NLRP1/ACE2 | a549-ascov2-nlrp1 | Invivogen |
| A549 ASC GFP ACE2 | a549-ascov2 | Invivogen |
| THP1 | thp-null | Invivogen |
| Recombinant DNA | Catalog reference | Provider |
| hNLRP1 gene | puno1-hnalp1a | Invivogen |
| . | . | . |
|---|---|---|
| Antibody | Catalog reference | Provider |
| Rabbit anti-ZAK antibody 1:1,000 | A301-993A | Bethyl Laboratories |
| P38 MAPK antibody 1:1,000 | 9212S | Cell Signaling |
| Phospho-P38 MAPK (Thr180/Tyr182) (D3F9) XPRabbit mAb 1:1,000 | 4511S | Cell Signaling |
| JNK1 Rabbit anti-human polyclonal 1:1,000 | 44690 G | Invitrogen |
| Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb 1:1,000 | 4668S | Cell Signaling |
| Anti-NLRP1 1:200 | 679802 | Biolegend |
| Anti-NLRP1 (N-terminal) 1:1,000 | AF6788-SP;RRID: AB_2916167 | R&D Systems |
| Anti-NLRP1 (C-terminal), 1:1,000 | ab36852;RRID: AB_776633 | Abcam |
| Anti-Sheep igG HRP (1/4,000) | HAF016;RRID: AB_562591 | R&D |
| Goat-anti-rabbit IgG (H+L), HRP conjugate (1/5,000) | R-05072-500;RRID: AB_10719218 | Advansta |
| GSDMDC1 antibody 1:1,000 | NBP2-33422 | Biotechne |
| Anti-puromycin antibody, clone 12D10 | MABE343 | Sigma-Aldrich |
| EEF2 antibody (1:1,000) | 2332S | Cell Signaling |
| DPH1 antibody (1:1,000) | #H00001801-M02 | Thermo Fisher Scientific |
| Anti-α Tubulin antibody 1:1,000 | ab4074 | Abcam |
| Streptavidin—HRP | 434323 | Thermo Fisher Scientific |
| Anti-NLRP3/NALP3,mAb (Cryo-2) | AG-20B-0014-C100 | AdipoGen |
| Reagents | Catalog reference | Provider |
| Transwell | 140620 | Thermo Fisher Scientific |
| NAD+, biotin-labeled | 80610 | BPSBioscience |
| Pseudomonas exotoxin A 0.5 mg | P0184-.5 MG | Sigma |
| Inactive exotoxin A | N.A. | Creative Diagnostics |
| Diphtheria toxin | D0564-1MG | Sigma-Aldrich |
| Cholix toxin | This study | This study |
| Anisomycin | SE-S7409-10 MG | Selleck |
| Talabostat (Val-boro-pro, VbP) | tlrl-VbP-10 | Invivogen |
| PhoSTOP | 4906845001 | Sigma-Aldrich/Roche |
| cOmplete, Mini, EDTA-free protease inhibitor cocktail | 4693159001 | Sigma-Aldrich/Roche |
| Lipofectamine LTX | 15338030 | Invitrogen |
| Puromycin dihydrochloride | A1113803 | Gibco |
| Molecular probes SYTOX Green nucleic acid stain | S7020 | Thermo Fisher Scientific |
| Propidium iodide | P1304MP | Thermo Fisher Scientific |
| Nate | lyec-nate | Invivogen |
| Phos-tag Acrylamide | AAL-107 | Wako Chemicals |
| Manganese chloride(II) | 63535 | Sigma-Aldrich |
| Prestained protein size marker III | 230-02461 | Wako Chemicals |
| Ivacaftor (VX-770) 5 mg/10 µM | S1144 | Selleck |
| Tezacaftor (VX-661) 5 mg/10 µM | S7059 | Selleck |
| Elexacaftor (VX-445) 5 mg/5 µM | S8851 | Selleck |
| Critical commercial assays | Catalog reference | Provider |
| Human total IL-18 duo set | DY318-05 | R&D |
| Cyquant LDH | C20301 | Thermo Fisher Scientific |
| Human IL-1β | 88-7261-77 | Thermo Fisher Scientific |
| Q5 site-directed mutagenesis kit protocol | E0554 | NEB |
| Chemicals | Catalog reference | Provider |
| ZAK inhibitor (PLX-4720) 10 µM | HY-51424 | Med Chem Express |
| P38 inhibitor SB 203580 10 µM | HY-10256 | Med Chem Express |
| Caspase-1 inhibitor—Ac-YVAD-cmk 20 µM | inh-yvad | Invivogen |
| Z-VAD-FMK 20 µM | vad-tlrl: | Invivogen |
| Caspase-3/7 inhibitor ZDEVD-FMK 20 µM | S7312 | Selleck |
| Caspase-8 inhibitor - Z-IETD-FMK | inh-ietd | Invivogen |
| Bortezomib 1 µM | S1013 | Selleck |
| CHX | C4859-1Ml | Sigma-Aldrich |
| MCC950, 10 mg | Inh-mcc | Invivogen |
| hPNECs | Catalog reference | Provider |
| Tryple express enzyme (1X), no phenol red | 10718463 | Thermo Fisher Scientific |
| Nutragen type I collagen solution, 6 mg/ml (Bovine) | 5010-50 Ml | Advanced Biomatrix |
| Sputolysin | 560000-1SET | Sigma-Aldrich |
| PneumaCult-Ex plus medium | 05040 | Stem Cell |
| Costar 6.5 mm transwell, 0.4 µm pore polyester membrane inserts | 38024 | Stem Cell |
| hPCECs | Catalog reference | Provider |
| Human corneal epithelial cell growth medium | 221-500 | Tebu-bio |
| Human corneal epithelial cells | 630-05a | Tebu-bio |
| Cell lines | Catalog reference | Provider |
| A549 ASC GFP NLRP1 | a549-ascg-nlrp1 | Invivogen |
| A549 ASC GFP | a549-ascg | Invivogen |
| A549 ASC GFP NLRP1/ACE2 | a549-ascov2-nlrp1 | Invivogen |
| A549 ASC GFP ACE2 | a549-ascov2 | Invivogen |
| THP1 | thp-null | Invivogen |
| Recombinant DNA | Catalog reference | Provider |
| hNLRP1 gene | puno1-hnalp1a | Invivogen |