KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-HA | GenScript | Cat# A01244–100: RRID: AB_1289306 |
| Mouse monoclonal anti-Flag | GenScript | Cat# A00187–200; RRID: AB_1720813 |
| Mouse monoclonal anti-NLRP3(Cryo-2) | Adipogen | Cat# AG-20B-0014-C100; RRID: AB_2490202 |
| Rabbit monoclonal anti-NEK7 | Abcam | Cat# ab133514; RRID: AB_2877625 |
| Rabbit monoclonal anti-GSDMD | Abcam | Cat# ab209845; RRID: AB_2783550 |
| Rabbit monoclonal anti-RACK1 | Cell Signaling | Cat# 5432; RRID: AB_10705522 |
| Rabbit monoclonal anti-S6 Ribosomal Protein (RPS6) | Cell Signaling | Cat# 2217S; RRID: AB_331355 |
| Mouse monoclonal IgG2b isotype control | Cell signaling | Cat# 53484S; RRID: AB_2799435 |
| Goat polyclonal anti-IL-1β | R&D system | Cat# AF-401-NA; RRID: AB_416684 |
| Rat polyclonal anti-ASC | Dr. Gabriel Nunez | N/A |
| Rabbit polyclonal anti-Caspase-1 | Dr. Gabriel Nunez | N/A |
| THE™ beta Actin Antibody [HRP] | GenScript | Cat# A00730–200; RRID: AB_914100 |
| Bacterial and Virus Strains | ||
| Salmonella strain SL1344 | Dr. Denise Monack | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| High-capacity streptavidin agarose resin | ThermoFisher | Cat# 20359 |
| High-capacity streptavidin agarose resin | GenScript | Cat# L00353 |
| S-protein agarose beads | EMD Millipore | Cat# 69704 |
| Protein G beads | GenScript | Cat# L00209 |
| LPS (Escherichia coli 055:B5, Ultrapure) | InvivoGen | Cat# tlrl-pb5lps |
| LPS-SM (Salmonella Minnesota R595,Ultrapure) | InvivoGen | Cat# tlrl-smlps |
| poly(dA:dT)/lyovec | InvivoGen | Cat# tlrl-patc |
| ATP | Sigma | Cat# A2383 |
| Gramicidin | Sigma | Cat# G5002 |
| L-leucyl-L-leucine methyl ester (LLOMe) | Sigma | Cat# L7393 |
| Nigericin | EMD Millipore | Cat# 481990 |
| bis[sulfosuccinimidy] suberate (BS3) | ThermoFisher | Cat# 21580 |
| Silica | US Silica | Cat# Min-U-Sil 5 |
| C-1 | Tocris | Cat# 0543 |
| GF 109203X | Tocris | Cat# 0741 |
| Go 6983 | Tocris | Cat# 2285 |
| Lipofectamine LTX | ThermoFisher | Cat# 15338100 |
| Lipofectamine 2000 | ThermoFisher | Cat# 11668019 |
| DOTAP | Biontex | Cat# T010–2.0 |
| Lenti-X concentrator | Clontech | Cat# 631232 |
| SYBR Premix ExTaq | Takara | Cat# RR820B |
| jetPEI solution | Polyplus Transfection | Cat# 101–10N |
| Critical Commercial Assays | ||
| Mouse TNF-α ELISA Kit | R&D Systems | Cat# DY410–05 |
| Mouse IL-1β ELISA Kit | R&D System | Cat# DY401–05 |
| LDH Cytotoxicity Detection Kit | TakaRa Bio | Cat# MK401 |
| QuikChange II XL Site-Directed Mutagenesis Kit | Agilent | Cat# 20051 |
| Cell line Nucleofector kit V | Lonza | Cat# 90279050 |
| ON-TARGETplus RACK1 siRNA pool | Darmacon | Cat# L-062125-00-0005 |
| ON-TARGETplus Non-targeting pool | Darmacon | Cat# D-001810-10-20 |
| RNeasy Mini kit | QIAGEN | Cat# 74104 |
| iScriptTM cDNA Synthesis kit | BioRad | Cat# 1708890 |
| KiCqStart Primers | Sigma, | Cat# KSPQ12012 |
| Experimental Models: Cell Lines | ||
| Immortalized bone marrow-derived macrophages (iBMDM) | (He et al., 2016b) | N/A |
| Nlrp3−/− iBMDM | (He et al., 2016b) | N/A |
| NEK7 KO iBMDM | (He et al., 2016b) | N/A |
| NLRP3 R258W iBMDM | Dr. Gabriel Nunez | N/A |
| HEK293T | ATCC | Cat# CRL-3216 |
| Experimental Models: Organisms/Strains | ||
| Mouse: C57BL/6J | Jackson Laboratories | Cat# 000664 |
| Oligonucleotides | ||
| Mouse RACK1 siRNA for in vivo: 5’ GUAGAUGAAUUGAAGCAAG 3’ | Biosynthesis | N/A |
| Control siRNA for in vivo: 5’ CUUGCUUCAAUUCAUCUAC 3’ | Biosynthesis | N/A |
| Human RACK1 siRNA: 5’ CGAUUUGUGGGCCAUACCA 3’ | Sigma | N/A |
| Control siRNA | Sigma | Cat# SIC001 |
| Recombinant DNA | ||
| RACK1 cDNA plasmid (pEGFP-N1-RACK1) | Addgene | Cat# 41088 |
| pHIV-EGFP | Addgene | Cat# 21373 |
| pHIV-NLRP3-SFP | (He et al., 2016b) | N/A |
| pHIV-NEK7-SFP | This paper | N/A |
| pHIV-ASC-SFP | This paper | N/A |
| pHIV-RACK1-HA | This paper | N/A |
| pHIV-RACK1R36D/K38E-HA | This paper | N/A |
| pCDNA3-NLRP3-Flag | (He et al., 2014) | N/A |
| pCDNA3-NLRP3-Δpyrin -Flag | (He et al., 2014) | N/A |
| pCDNA3-NLRP3-ΔLRR -Flag | (He et al., 2014) | N/A |
| pCDNA3-NLRP3-LRR-Flag | (He et al., 2014) | N/A |
| pCDNA3-NLRP3-NOD-Flag | (He et al., 2014) | N/A |
| pCDNA3-NLRP3-pyrin-Flag | (He et al., 2014) | N/A |
| pLKO.1-shRACK1 #1 | Sigma | TRCN0000287170 |
| pLKO.1-shRACK1 #2 | Sigma | TRCN0000294547 |
| pCMV-dR8.2 dvpr | Addgene | Cat# 8455 |
| pCMV-VSV-G | Addgene | Cat# 8454 |
| pLKO scramble | Addgene | Cat# 1864 |
| pcDNA3.1- YFP-NLRP3-Luc | (Tapia-Abellán et al., 2019) | N/A |
| Software and Algorithms | ||
| ImageJ | https://imagej.nih.gov | N/A |
| GraphPad Prism 7.0 | GraphPad Software | N/A |
| Other | ||
| 8-well permanox chamber slide | Thermo Scientific | Cat# 177445 |
| PVDF membrane | EMD Millipore | Cat# IPVH00010 |