Table 2.
Reagents and resources.
| Name | Source | Identifier |
|---|---|---|
| Antibodies | ||
| anti-TRIF | Proteintech | Cat#23288-1-AP |
| anti-pIRF3 (Ser396) | BIOSS | Cat#bs-3195R |
| anti-STAT1 | Abcam | Cat#ab99415 |
| anti-pSTAT1 (pY701) [M135] | Abcam | Cat#ab29045 |
| anti-GBP5 | Proteintech | Cat#13220-1-AP |
| anti-GBP2 | Proteintech | Cat#11854-1-AP |
| anti-Caspase 11 (17D9) | eBioscience™ | Cat#14-9935-82 |
| anti-Caspase-11 | Abcam | Cat# ab180673 |
| anti-CASPASE-4 | MBL | Cat#M029-3 |
| anti-GSDMD | Proteintech | Cat#20770-1-AP |
| anti-Caspase-1 | AdipoGen Life Sciences | Cat#AG-20B-0042-C100 |
| anti-Caspase-1 (14F468) | Santa Cruz Biotechnology | Cat#sc-56036 |
| anti-IL-1β/IL-1F2 | R&D Systems | Cat#AF-401-NA |
| anti-IL-1β/IL-1F2 | R&D Systems | Cat#MAB4011 |
| anti-IL-1β (B122) | Santa Cruz Biotechnology | Cat#sc-12742 |
| anti-Caspase-8 (1G12) | Enzo Life Sciences | Cat#ALX-804-447-C100 |
| anti-β-actin | Cell Signaling Technology | Cat#4967s |
| anti-β-actin | ThermoFisher Scientific | Cat#PA1-183 |
| anti-β-actin | R&D Systems | Cat#MAB-8929 |
| Mouse anti-armenian hamster IgG-HRP | Santa Cruz Biotechnology | Cat#sc-2789 |
| Goat anti-rabbit IgG(H+L), HRP conjugate | Proteintech | Cat#SA00001-2 |
| IRDye® 800CW goat anti-rabbit IgG | LI-COR Biosciences | Cat#926-32211 |
| IRDye® 680RD goat anti-rabbit IgG | LI-COR Biosciences | Cat#926-68071 |
| Goat anti-mouse IgG(H+L), HRP conjugate | Proteintech | Cat#SA00001-1 |
| IRDye® 680RD goat anti-mouse IgG | LI-COR Biosciences | Cat#926-68070 |
| Mouse IgG (H&L) secondary antibody peroxidase conjugated pre-adsorbed | Rockland Immunochemicals | Cat#610-1319-0500 |
| Rabbit anti-goat IgG(H+L), HRP conjugate | Proteintech | Cat#SA00001-4 |
| Goat anti-Rat IgG(H+L), HRP conjugate | Proteintech | Cat#SA00001-15 |
| IRDye® 800CW goat anti-rat IgG | LI-COR Biosciences | Cat#926-32219 |
| anti-IFNAR1 (Clone: MAR1-5A3) | BioLegend | Cat#127302 |
| anti-IgG1 (Clone: MOPC-21) | BioLegend | Cat#400102 |
| Bacterial Strains | ||
| Escherichia coli (K12) | Dr. Jason Gardner (gardnejr@ucmail.uc.edu) |
N/A |
| Klebsiella pneumoniae (KP2 2-70) | Dr. Jason Gardner (gardnejr@ucmail.uc.edu) |
N/A |
| Group A Streptococcus (Streptococcus pyogenes) | Dr. Suba Nookala (suba.nookala@und.edu) | N/A |
| Primers | ||
| Mt3 | Applied Biosystems | Mm00496661_g1 |
| Mt2 | Applied Biosystems | Mm00809556_s1 |
| Mt1 | Applied Biosystems | Mm00496660_g1 |
| Hprt | Applied Biosystems | Mm00446968_m1 |
| MT3 | Applied Biosystems | Hs00359394_g1 |
| MT2A | Applied Biosystems | HS02379661_g1 |
| HPRT1 | Applied Biosystems | Hs99999909_m1 |
| Genotyping Primers | IDT | N/A |
| siRNA and expression vector | ||
| MT3 | Ambion | Cat#AM16708 |
| Ticam1 | Dharmacon™ | Cat#L-055987-00-0005 |
| ON-TARGETplus™ Control Pool (Non-Targeting pool) | Dharmacon™ | Cat#D-001810-10-20 |
| pCMV6-AC-GFP (PS100010) | ORIGENE | Cat#MG200059 |
| pCMV6-AC-GFP | ORIGENE | Cat# PS100010 |
| Chemicals and accessories | ||
| RPMI 1640 | CORNING | REF#10-041-CV |
| Opti-MEM® I | GIBCO | REF#31985-070 |
| FBS | HyClone | Cat#SH30396.03 |
| DPBS | CORNING | REF#21-031-CV |
| PBS without calcium and magnesium | CORNING | REF#21-040-CV |
| HBSS | CORNING | REF#21-021-CM |
| HEPES | Sigma | Cat#H3375 |
| Mouse M-CSF | PEPROTECH | Cat#315-02 |
| Human M-CSF | PEPROTECH | Cat#300-25 |
| Trypsin-EDTA | CORNING | REF#25-053-CI |
| LPS-B5 Ultrapure | InVivoGen | Cat#tlrl-pb5lps |
| Glycerol | Fisher Bioreagents | Cat#BP229-1 |
| LB Broth | Fisher Bioreagents | Cat#BP1427-500 |
| BBL™ Brain Heart Infusion Broth | Becton Dickinson | Cat#22182 |
| Agar | BD Bacto™ | Cat#214010 |
| BD Bacto™ Dehydrated Culture Media: Todd Hewitt Broth | BD | Cat#249240 |
| Gibco™ Bacto™ Yeast Extract | Gibco | Cat#212750 |
| BD BBL™ RODAC™ Trypticase™ Soy Agar with 5% Sheep Blood (TSA II) | BD | Cat#221261 |
| EZ Pack™ Agarose | ASi | Item No.#AG2501 |
| MCC950 | ApexBio | B7946-50 |
| ZnSO4 | Sigma | Cat#z-4750 |
| Sodium Chloride | Sigma | Cat#S1679-1KG |
| Ethyl Alcohol | Fisher Scientific | Cat#A407-4 |
| Methanol | Fisher Scientific | Cat#A434-20 |
| Methanol | BDH | Cat#BDH1135-4LP |
| Dimethyl Sulfoxide | Sigma | Cat#D8418-1L |
| Chloroform | Fisher Scientific | Cat#C5312 |
| Hydrochloric Acid | Fisher Scientific | Cat#A508-212 |
| Sulfuric Acid | Fisher Scientific | Cat#A300SI-212 |
| Sodium Hydroxide | Fisher Scientific | Cat#S318-3 |
| Precise Protein Gels | Invitrogen | REF#XP04205BOX |
| SurePAGE™, Bis-Tris, 10x8 gels (4-20%, 15 wells) | GenScript | Cat#M00657 |
| Tris-MOPS-SDS Running Buffer Powder | GenScript | Cat#M00138 |
| Nitrocellulose membranes | BIO-RAD | Cat#162-0112 |
| Protease &Phosphatase Inhibitor Cocktail | Thermo Scientific | Cat#78442 |
| Denaturing Cell Extraction Buffer | Invitrogen | Cat#FNN0091 |
| XCell II™ Blot Module | ThermoFisher Scientifics | Cat#EI9051 |
| Glycine | Fisher Scientific | Cat#BP381 |
| Tris-base | Fisher Scientific | Cat#BP152 |
| SDS | Fisher Scientific | Cat#BP166 |
| Tween 20 | Acros Organics | Cat#23336-0010 |
| Tween 20 | Sigma | Cat#P-1379 |
| Triton X100 | Fisher Scientific | Cat#BP151 |
| Bovine serum albumin | Sigma | Cat#A7030 |
| Intercept® (TBS) Blocking Buffer | LI-COR Biosciences | Cat#927-60001 |
| Non-fat dry milk | Nash Finch Co. | N/A |
| Isoflurane | Covetrus | NDC Code(s)#11695-6777-2 |
| Nair | Chuech & Dwight Co.inc | N/A |
| Gauze | Medline | REF#PRM25444 |
| Puritan cotton tipped applicator | Puritan | SKU#836-WC |
| Insulin injection syringe (1mL) | Exelint international co. | Cat#26029 |
| 10 ml plastic Syringe | Fisherbrand | Cat#14955459 |
| 1 ml plastic Syringe | Fisherbrand | Cat#14955456 |
| BioLite12 well multidish | Thermo Scientific | Lot# H4XA4RE106 |
| 24 well tissue culture plates | CellPro | Lot# 072219BA03 |
| 48 well cell culture plate | NEST | Lot# 121717A004 |
| Tissue culture plate 96 well, flat bottom | Fisherbrand | Cat#FB012931 |
| 96 well ELISA plate | NEST | Lot#04291818A007 |
| Petri dish | NEST | Lot#753001 |
| 15 ml Centrifuge tube | Fisher Scientific | Cat#14-955-237 |
| 50 ml Centrifuge tube | Fisher Scientific | Cat#14-955-239 |
| Cell scraper | SPL Life Sciences | Cat#90030 |
| Golden Rod animal Lancet (4mm) | Medipoint Inc | N/A |
| Omni homogenizer Th-01 and tips | Omni International | Model Number#THP115 |
| GeneArt Platinum Cas9 Nuclease Alt-R SpCas9 Nuclease 3NLS sgRNA |
ThermoFisher ThermoFisher CCHMC transgenic core |
Cat# B25641 Cat# 1074181 N/A |
| TSK gel 3000SW gel filtration column | TOSOH BIOSCIENCE | Cat#05789 |
| DreamTaq DNA Polymerase | Thermo Scientific | REF#EP0702 |
| Commercial Assays | ||
| TransIT®-TKO | Mirus Bio LLC | Prod. No.#MIR 2150 |
| TransIT®-LT1 | Mirus Bio LLC | Prod. No.#MIR 2300 |
| Pierce Protein Transfection Reagent Kit | Thermo Scientific | REF#89850 |
| Human IL-1β/IL-1F2 DuoSet ELISA | R&D Systems | Cat#DY201-05 |
| Mouse IL-1β | BioLegend | Cat# 432601 |
| IL-1α | BioLegend | Cat# 433401 |
| RNeasy Mini Kit | Qiagen | Cat# 74104 |
| QUICK-RNA™ MINIPREP KIT | Denville Scientific | Cat# R1055 |
| Reverse Transcription System | Promega | REF#A3500 |
| Probe Lo-Rox 2X qPCR Mix | RADIANT™ | Cat#QP9020 |
| CytoTox 96® NonRadioactive Cytotoxicity Assay kit Promega | Promega | REF#G1781 |
| SuperSignal™ West Femto Maximum Sensitivity Substrate | Thermo Scientific | Cat#34096 |
| NEBNext Poly(A) mRNA Magnetic Isolation Module | New England BioLabs | Cat#E7490L |
| NEBNext Ultra Directional RNA Library Prep Kit | New England BioLabs | Cat# E7420L |
| NEBNext Library Quant Kit | New England BioLabs | Cat# E7630L |
| MEGAshorscript T7 Kit | Thermo Fisher | Cat#AM1354 |
| MEGAclear Kit | Thermo Fisher | Cat# AM1908 |
| Quick-DNA™ Miniprep Plus Kit | ZYMO RESEARCH | Cat#D4069 |
| Radiant™ Taq DNA Polymerase | RADIANT™ | Cat#C101 |
| Experimental Models: Organisms/Strains | ||
| C57BL/6 Mice | Jackson Laboratory | Stock#00064 |
| C57BL/6 Mt3-/- deletion of Exon 3 | Transgenic Animal and Genome editing core facility at CCHMC | N/A |
| C57BL/6 Mt1-/-Mt2-/- | Dr. George S. Deepe (deepegs@ucmail.uc.edu) | N/A |
| C57BL/6 Casp4tm1Yuan /J (Casp-11-/- ) | Jackson Laboratory | Stock#024698 |
| Mt3-/-Casp-11-/- | Crossed and bred in-house | N/A |
| C57BL/6 Lys2Cre | Dr. George S. Deepe (deepegs@ucmail.uc.edu) | N/A |
| C57BL/6 Lys2Cre Mt3fl/fl | Crossed and bred in-house | N/A |
| Instrument, Software and Algorithms | ||
| FluorChem® HD2 | Cell Biosciences | S/N#FC HD2 Imager |
| Odyssey CLx Imaging system | LI-COR | N/A |
| 7500 Fast Real Time PCR System | Applied Biosystems | S/N#275013253 |
| RNA-seq data analysis - DAVID Bioinformatics Resources v.6.8 | NIAID/NIH | (https://david.ncifcrf.gov/summary.jsp) |
| Protein-protein interaction networks- STRING v.11.0 | ELIXIR | (https://string-db.org/) |
| NIH ImageJ Fiji | NIH | (https://imagej.nih.gov/ij/) |