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. 2023 Aug 14;42(19):e113481. doi: 10.15252/embj.2023113481
Reagent/resource Reference or source Identifier or catalog number
Experimental models
HEK293T ATCC Cat# ACS‐4500
THP‐1 ATCC Cat# TIB‐202
L929 Je‐Wook Yoo N/A
iBMDM Je‐Wook Yoo N/A
iBMDM NLRP3‐GFP Je‐Wook Yoo N/A
Citrobacter rodentium ATCC Cat# 51459
Salmonella typhimurium ATCC Cat# 14028
Pseudomonas aeruginosa ATCC Cat# BAA‐1744
March5 fl/fl mice This paper N/A
March5 fl/fl;Lyz‐cre mice This paper N/A
Recombinant DNA
pcDNA3.1 Invitrogen Thermo Fisher Cat# V79020
pcDNA3.1‐myc‐MARCH5 This paper N/A
pcDNA3.1‐myc‐MARCH5 (2KR) This paper N/A
pcDNA3.1‐myc‐MARCH5 (H43W) This paper N/A
pcDNA6‐SFB‐MARCH5 WT This paper N/A
pcDNA6‐SFB‐MARCH5 (2KR) This paper N/A
pCMV6‐AC‐cMyc‐DDK This paper N/A
pCMV6‐NLRP3 WT‐MycDDK This paper N/A
pCMV6‐NLRP3 K93R‐MycDDK This paper N/A
pCMV6‐NLRP3 K192R‐MycDDK This paper N/A
pCMV6‐NLRP3 K194R‐MycDDK This paper N/A
pCMV6‐NLRP3 K324R‐MycDDK This paper N/A
pCMV6‐NLRP3 K430R‐MycDDK This paper N/A
pCMV6‐NLRP3 K689R‐MycDDK This paper N/A
pCMV6‐NLRP3 K696R‐MycDDK This paper N/A
pCMV6‐NLRP3 S194A‐MycDDK This paper N/A
pcDNA3.1/Myc‐NLRP3 WT Ren et al 2019 N/A
pcDNA3.1/Myc‐NLRP3 ▵PYD Ren et al 2019 N/A
pcDNA3.1/Myc‐NLRP3 ▵Linker Ren et al 2019 N/A
pcDNA3.1/Myc‐NLRP3 ▵NACHT Ren et al 2019 N/A
pcDNA3.1/Myc‐NLRP3 ▵LRR Ren et al 2019 N/A
pCMV3‐C‐FLAG‐IL‐1β Sino Biological Cat# HG10139‐CF
pCMV3‐C‐HA‐NEK7 Sino Biological Cat# HG11534‐CY
Antibodies
Mouse monoclonal anti‐NLRP3 (Cryo2‐)

Adipogen

1:5,000

Cat# AG‐20B‐0014
Rabbit polyclonal anti‐NLRP3

Cell Signaling Technology

1:1,000

Cat# 15101
Goat polyclonal anti‐NLRP3

Abcam

1:1,000

Cat# ab4207
Rabbit polyclonal anti‐ASC

Adipogen

1:1,000

Cat# AG‐25B‐0006
Mouse monoclonal anti‐ASC

Santa Cruz Biotechnology

1:1,000

Cat# sc‐514414
Rabbit polyclonal anti‐ASC

Santa Cruz Biotechnology

1:1,000

Cat# sc‐22514‐R
Mouse monoclonal anti‐IL‐1β

Cell Signaling Technology

1:1,000

Cat# 12242
Goat Polyclonal anti‐IL‐1β (p20)

Adipogen

1:1,000

Cat# AF‐401‐NA
Mouse monoclonal anti‐IL‐1β

Santa Cruz Biotechnology

1:1,000

Cat# 12742
Mouse monoclonal anti‐mouse caspase‐1

Adipogen

1:1,000

Cat# AG‐20B‐0042
Rabbit polyclonal anti‐human caspase‐1

Cell Signaling Technology

1:1,000

Cat# 3866
Mouse monoclonal anti‐human caspase‐1

Santa Cruz Biotechnology

1:1,000

Cat# sc‐56036
Mouse monoclonal anti‐mouse MAVS

Cell Signaling Technology

1:1,000

Cat# 83000
Rabbit polyclonal anti‐human MAVS

Cell Signaling Technology

1:1,000

Cat# 3993
Rabbit polyclonal anti‐MARCH5

Shigeru YANAGI

1:1,000

N/A
Mouse monoclonal anti‐NEK7

Santa Cruz Biotechnology

1:1,000

Cat# sc‐393539
Rabbit monoclonal anti‐NEK7

Abcam

1:1,000

Cat# ab133514
Mouse monoclonal anti‐FLAG

Sigma‐Aldrich

1:1,000

Cat# F1804
Mouse monoclonal anti‐c‐Myc

Santa Cruz Biotechnology

1:1,000

Cat# sc‐40
Mouse monoclonal anti‐HA

Santa Cruz Biotechnology

1:1,000

Cat# sc‐7392
Mouse monocclonal anti‐Tubulin

Santa Cruz Biotechnology

1:1,000

Cat# sc‐73242
Rabbit polyclonal anti‐GAPDH

Cell Signaling Technology

1:1,000

Cat# 5174
Goat anti Mouse IgG (H+L)‐HRP conjugate

Bio‐rad

1:5,000

Cat# 170516
Goat anti Rabbit IgG (H+L)‐HRP conjugate

Bio‐rad

1:5,000

Cat# 1706515
Donkey anti Goat IgG (H+L)‐HRP conjugate

Bio‐rad

1:5,000

Cat# 642005
Oligonucleotides and other sequence‐based reagents

Human MARCH5 targeted siRNA

5′‐GG GUG GAA UUG CGU UUG UU‐3′

Bioneer This paper

Mouse March5 targeted siRNA

5′‐GGU UGU AGG CCA UAA AGA A‐3′

Bioneer This paper

Human MAVS targeted siRNA

5′‐CAG GUU GGC CUC AUG AGA U‐3′

Bioneer Cat# 57506

Mouse Asc targeted siRNA

5′‐GCU CUU CAG UUU CAC ACC A‐3′

Bioneer This paper
Chemicals, Enzymes and other reagents
Lipopolysaccharides from Escherichia coli O26:B6 Sigma‐Aldrich Cat# L8274
ATP Sigma‐Aldrich Cat# A6419
Nigericin Invivogen Cat# tlrl‐nig
FLA‐ST Invivogen Cat# tlrl‐stfla
Poly(I:C) Invivogen Cat# tlrl‐pic
Poly(dA:dT) Invivogen Cat# tlrl‐patn‐1
Recombinant Human MARCH5 GST Protein Abnova Cat# H00054708‐P0
Anti‐FLAG M2 affinity Gel Sigma‐Aldrich Cat# A2220
DSS (disuccinimidyl suberate) Invitrogen Thermo Fisher Cat# A39267
Software
GraphPad Prism 8.0 GraphPad, San Diego, CA https://www.graphpad.com/scientific‐software/prism/
ImageJ National Institues of Health https://imagej.nih.gov/nih‐image/
Biorender N/A https://biorender.com/
Ubpred N/A http://ubpred.org/
Other
LDH‐Glo™ Cytotoxicity Assay Promega Cat# J2380
Caspase‐Glo® 1 Inflammasome Assay Promega Cat# G9951l
Lipofectamine 2000 Invitrogen Cat# 11668‐019
ELISA MAX™ Deluxe Set Mouse IL‐1β Biolegend Cat# 432616
Mouse Caspase‐1 ELISA Kit Novus biologicals Cat# NBP2‐75014
BD OptEIA™ Mouse TNF (Mono/Mono) ELISA Set BD OptEIA Cat# 555268
BD OptEIA™ Mouse IL‐6 ELISA Set BD OptEIA Cat# 555240
IL‐1 beta Human Uncoated ELISA Kit Invitrogen Cat# 88‐7261‐22
Recombinant Mouse IL‐18 ELISA Set MBL Cat# B002‐5