wizard cryo 1 crystallization screen Search Results


piezoelectric stacks part no. pst150/5x5/7 cryo 1  (Piezomechanik GmbH)
90
Piezomechanik GmbH piezoelectric stacks part no. pst150/5x5/7 cryo 1
Piezoelectric Stacks Part No. Pst150/5x5/7 Cryo 1, supplied by Piezomechanik GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/piezoelectric+actuator+pst150+5+60/pmc08866430-121-10-18
Average 90 stars, based on 1 article reviews
piezoelectric stacks part no. pst150/5x5/7 cryo 1 - by Bioz Stars, 2026-09
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cryo 1  (Emerald BioStructures Inc)
90
Emerald BioStructures Inc cryo 1
Cryo 1, supplied by Emerald BioStructures Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/cryo+1/pmc03339967-260-8-12
Average 90 stars, based on 1 article reviews
cryo 1 - by Bioz Stars, 2026-09
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anti-nlrp3  (Enzo Biochem)
90
Enzo Biochem anti-nlrp3
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Anti Nlrp3, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/nlrp3+antibody/pmc04855944-554-10-11
Average 90 stars, based on 1 article reviews
anti-nlrp3 - by Bioz Stars, 2026-09
90/100 stars
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cryoprobe 3011 linear  (AtriCure Inc)
90
AtriCure Inc cryoprobe 3011 linear
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Cryoprobe 3011 Linear, supplied by AtriCure Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/cryo1+probe/pm23422798-10-30-38
Average 90 stars, based on 1 article reviews
cryoprobe 3011 linear - by Bioz Stars, 2026-09
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freezing container  (Thermo Fisher)
99
Thermo Fisher freezing container
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Freezing Container, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/Mr%2E+Frosty+Freezing+Container/pm19057140-56-10-12
Average 99 stars, based on 1 article reviews
freezing container - by Bioz Stars, 2026-09
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cryo 1  (Jena Bioscience)
94
Jena Bioscience cryo 1
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Cryo 1, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/JBScreen+Classic+1/pmc04188077-71-24-32
Average 94 stars, based on 1 article reviews
cryo 1 - by Bioz Stars, 2026-09
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cryo 1  (Emerald BioSystems Inc)
90
Emerald BioSystems Inc cryo 1
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Cryo 1, supplied by Emerald BioSystems Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/cryo+1/pmc02720352-84-6-8
Average 90 stars, based on 1 article reviews
cryo 1 - by Bioz Stars, 2026-09
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container  (Thermo Fisher)
99
Thermo Fisher container
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Container, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/Minocycline+hydrochloride/pm15153480-144-10-19
Average 99 stars, based on 1 article reviews
container - by Bioz Stars, 2026-09
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piezomechanick 5x5x9 pzt piezoelectric stack  (Piezomechanik GmbH)
90
Piezomechanik GmbH piezomechanick 5x5x9 pzt piezoelectric stack
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Piezomechanick 5x5x9 Pzt Piezoelectric Stack, supplied by Piezomechanik GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/piezomechanick+5x5x9+pzt+piezoelectric+stack/pmc06462099__pnas__1818910116__sapp-74-24-27
Average 90 stars, based on 1 article reviews
piezomechanick 5x5x9 pzt piezoelectric stack - by Bioz Stars, 2026-09
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cryo 1◦c freezing container  (Thermo Fisher)
99
Thermo Fisher cryo 1◦c freezing container
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Cryo 1◦C Freezing Container, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/Trypan+Blue/pm18432951-16-1-5
Average 99 stars, based on 1 article reviews
cryo 1◦c freezing container - by Bioz Stars, 2026-09
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carbotrap /glass beads  (Tekmar Inc)
90
Tekmar Inc carbotrap /glass beads
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Carbotrap /Glass Beads, supplied by Tekmar Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/carbotrap++glass+beads/10__1016_slash_s0021___9673_ascii40_97_ascii41_00741___3-46-41-74
Average 90 stars, based on 1 article reviews
carbotrap /glass beads - by Bioz Stars, 2026-09
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cryotubes  (Thermo Fisher)
94
Thermo Fisher cryotubes
PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, <t>NLRP3,</t> ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).
Cryotubes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/wizard+cryo+1+crystallization+screen/Nunc+CryoTube+Colored+Caps/pm21948163-182-50-53
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cryotubes - by Bioz Stars, 2026-09
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Image Search Results


PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, NLRP3, ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: PTPN22 knockdown was induced in THP-1 cells using lentiviral shRNA expression vectors. Cells were pretreated for 12 hours with upLPS before activation with MDP (100 ng/ml, 24 hours), MSU (150 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 12 hours), or ATP (200 mM, 30 minutes). (A) Cell culture supernatants were analyzed for IL-1β secretion, LDH release, and IL-6 secretion by ELISA. (B) Cell lysates or supernatants were analyzed for caspase-1 (Casp-1), caspase-3 (Casp-3), IL-1β, NLRP3, ASC, and PTPN22 expression. Blots for IL-1β were run on the same gel but were discontinuous. Data are representative of 1 of at least 3 independent experiments with 3–5 replicas (n = 3–5; *P < 0.05, **P < 0.01; Newman-Keuls post hoc test). Numbers below the blots show results of densitometry (cleaved forms).

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: shRNA, Expressing, Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

THP-1 cells, MM6 cells, and BMDCs were treated with upLPS for 12 hours prior to activation with MDP (100 ng/ml, 24 hours), MSU (100 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 24 hours), ATP (2 mM, 30 minutes), dsDNA, or flagellin, as indicated. (A and C) PTPN22 was precipitated from THP-1 or MM6 lysates and analyzed by Western blot for coprecipitated NLRP3. (B) PTPN22 was precipitated from BMDCs and analyzed for NLRP3. (D) PTPN22 was precipitated from dsDNA-treated THP-1 cells or BMDCs and analyzed by Western blot for coprecipitation of AIM2. (E) PTPN22 was precipitated from dsDNA-treated THP-1 cells or BMDCs and analyzed by Western blot for coprecipitation of NLRC4. Data are representative of 1 of 3–5 independent experiments.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: THP-1 cells, MM6 cells, and BMDCs were treated with upLPS for 12 hours prior to activation with MDP (100 ng/ml, 24 hours), MSU (100 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 24 hours), ATP (2 mM, 30 minutes), dsDNA, or flagellin, as indicated. (A and C) PTPN22 was precipitated from THP-1 or MM6 lysates and analyzed by Western blot for coprecipitated NLRP3. (B) PTPN22 was precipitated from BMDCs and analyzed for NLRP3. (D) PTPN22 was precipitated from dsDNA-treated THP-1 cells or BMDCs and analyzed by Western blot for coprecipitation of AIM2. (E) PTPN22 was precipitated from dsDNA-treated THP-1 cells or BMDCs and analyzed by Western blot for coprecipitation of NLRC4. Data are representative of 1 of 3–5 independent experiments.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Activation Assay, Western Blot

THP-1 and MM6 cells (A and C), BMDCs (B), and PBMCs (D) were treated with upLPS for 12 hours prior to activation with MDP (100 ng/ml, 24 hours), MSU (100 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 24 hours), ATP (2 mM, 30 minutes), dsDNA, or flagellin, as indicated. NLRP3 was precipitated from cell lysates, and precipitates were analyzed for tyrosine phosphorylation, coprecipitated PTPN22, or coprecipitated PTPN2. (D) PTPN22 was precipitated in addition to NLRP3 and analyzed for coprecipitated NLRP3. (E) Amounts of NLRP3 and pTyr in NLRP3 precipitates were quantified using standard curves for NLRP3 and a pTyr peptide. The graph below the representative Western blots shows statistical analysis of densitometry. ND, not detected. Left and right blots in E were run on the same gel, but were discontinuous. Numbers below the blots in D show the amount of loaded NLRP3 or pTyr peptide (left) and measured amounts (right), respectively. The pound symbol (#) marks blots where the 130-kDa and 120-kDa forms of NLRP3 are not completely separated and appear as single bands. Data shown are from 1 of 3–5 independent experiments. *P < 0.05, **P < 0.01, Student’s t test with Bonferroni correction.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: THP-1 and MM6 cells (A and C), BMDCs (B), and PBMCs (D) were treated with upLPS for 12 hours prior to activation with MDP (100 ng/ml, 24 hours), MSU (100 ng/ml, 6 hours), TiO2 (150 ng/ml, 24 hours), SiO2 (150 ng/ml, 24 hours), ATP (2 mM, 30 minutes), dsDNA, or flagellin, as indicated. NLRP3 was precipitated from cell lysates, and precipitates were analyzed for tyrosine phosphorylation, coprecipitated PTPN22, or coprecipitated PTPN2. (D) PTPN22 was precipitated in addition to NLRP3 and analyzed for coprecipitated NLRP3. (E) Amounts of NLRP3 and pTyr in NLRP3 precipitates were quantified using standard curves for NLRP3 and a pTyr peptide. The graph below the representative Western blots shows statistical analysis of densitometry. ND, not detected. Left and right blots in E were run on the same gel, but were discontinuous. Numbers below the blots in D show the amount of loaded NLRP3 or pTyr peptide (left) and measured amounts (right), respectively. The pound symbol (#) marks blots where the 130-kDa and 120-kDa forms of NLRP3 are not completely separated and appear as single bands. Data shown are from 1 of 3–5 independent experiments. *P < 0.05, **P < 0.01, Student’s t test with Bonferroni correction.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Activation Assay, Western Blot

(A) THP-1 cells expressing control or PTPN22-targeting shRNA were treated with control or NLRP3-targeting siRNA constructs and incubated for 12 hours with upLPS before activation with MSU (6 hours). NLRP3 or PTPN22 was precipitated from the cell lysates and analyzed for tyrosine phosphorylation and PTPN22 coprecipitation or NLRP3 coprecipitation, respectively. (B) Purified NLRP3 was incubated in phosphatase buffer for 60 minutes in the presence or absence of purified WT-PTPN22 (WT), altered-function PTPN22 (619W), loss-of-function PTPN22 (263Q), or purified PTPN2. (C) Multiple sequence alignment of NLRP3 sequences from the indicated species using FASTA.2 (http://www.ebi.ac.uk) reveals conservation of the tyrosine at position 861. (D) HEK293T cells were transfected with WT NLRP3 or a NLRP3 construct where Tyr861 was replaced by a phenylalanine (Y>F). NLRP3 was immunoprecipitated and analyzed for the presence of pTyr. Lys, lysates. (E and F) Nlrp3–/– BMDCs were left nontransfected or transfected with WT NLRP3; Y>F NLRP3; or NLRP3 where Tyr858 was replaced with a glutamine to mimic constitutive phosphorylation (Y>E) or by an alanine (control to glutamine substitution [Y>A]) and activated with MSU. Lysates (E) and supernatants (F) were analyzed for the indicated proteins. (G) BMDCs were derived from ASC-, caspase-1–, NLRP3-, or PTPN22-deficient mice or mice expressing the autoimmunity-associated PTPN22 variant (619W); pretreated for 12 hours with upLPS; and activated with MSU. NLRP3 was immunoprecipitated and analyzed for pTyr and PTPN22. All data are representative of 1 of 3–5 independent experiments. Numbers below the blots show results of densitometry.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: (A) THP-1 cells expressing control or PTPN22-targeting shRNA were treated with control or NLRP3-targeting siRNA constructs and incubated for 12 hours with upLPS before activation with MSU (6 hours). NLRP3 or PTPN22 was precipitated from the cell lysates and analyzed for tyrosine phosphorylation and PTPN22 coprecipitation or NLRP3 coprecipitation, respectively. (B) Purified NLRP3 was incubated in phosphatase buffer for 60 minutes in the presence or absence of purified WT-PTPN22 (WT), altered-function PTPN22 (619W), loss-of-function PTPN22 (263Q), or purified PTPN2. (C) Multiple sequence alignment of NLRP3 sequences from the indicated species using FASTA.2 (http://www.ebi.ac.uk) reveals conservation of the tyrosine at position 861. (D) HEK293T cells were transfected with WT NLRP3 or a NLRP3 construct where Tyr861 was replaced by a phenylalanine (Y>F). NLRP3 was immunoprecipitated and analyzed for the presence of pTyr. Lys, lysates. (E and F) Nlrp3–/– BMDCs were left nontransfected or transfected with WT NLRP3; Y>F NLRP3; or NLRP3 where Tyr858 was replaced with a glutamine to mimic constitutive phosphorylation (Y>E) or by an alanine (control to glutamine substitution [Y>A]) and activated with MSU. Lysates (E) and supernatants (F) were analyzed for the indicated proteins. (G) BMDCs were derived from ASC-, caspase-1–, NLRP3-, or PTPN22-deficient mice or mice expressing the autoimmunity-associated PTPN22 variant (619W); pretreated for 12 hours with upLPS; and activated with MSU. NLRP3 was immunoprecipitated and analyzed for pTyr and PTPN22. All data are representative of 1 of 3–5 independent experiments. Numbers below the blots show results of densitometry.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Expressing, shRNA, Construct, Incubation, Activation Assay, Purification, Sequencing, Transfection, Immunoprecipitation, Derivative Assay, Variant Assay

Acute colitis was induced in female WT, Ptpn22–/–, or Nlrp3–/– mice by administration of 2.5% DSS for 7 days. Immunohistochemistry of the distal colon stained for CD3+ T cells (A) and Gr1+ granulocytes (B). Statistical analysis of infiltrating cells is shown in C. Data are representative of 1 of 2 independent experiments with 4–6 mice per group each (n = 4–6). Each dot represents 1 mouse. *P < 0.05, Mann-Whitney U test with Bonferroni correction. Original magnification (IHC), ×10.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: Acute colitis was induced in female WT, Ptpn22–/–, or Nlrp3–/– mice by administration of 2.5% DSS for 7 days. Immunohistochemistry of the distal colon stained for CD3+ T cells (A) and Gr1+ granulocytes (B). Statistical analysis of infiltrating cells is shown in C. Data are representative of 1 of 2 independent experiments with 4–6 mice per group each (n = 4–6). Each dot represents 1 mouse. *P < 0.05, Mann-Whitney U test with Bonferroni correction. Original magnification (IHC), ×10.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Immunohistochemistry, Staining, MANN-WHITNEY

Acute colitis was induced in female WT, Ptpn22–/–, or Nlrp3–/– mice by administration of 2.5% DSS for 7 days. (A) Weight development, (B) MPO activity, and (C) colon length. (D) H&E staining from the distal colon and (E) analysis of epithelial damage and infiltration. All data are representative of 1 of 2 independent experiments with 4–6 mice per group each (n = 4–6). Each dot represents 1 mouse. *P < 0.05, **P < 0.01, Mann-Whitney U test with Bonferroni correction. Original magnification (H&E and IHC), ×10.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: Acute colitis was induced in female WT, Ptpn22–/–, or Nlrp3–/– mice by administration of 2.5% DSS for 7 days. (A) Weight development, (B) MPO activity, and (C) colon length. (D) H&E staining from the distal colon and (E) analysis of epithelial damage and infiltration. All data are representative of 1 of 2 independent experiments with 4–6 mice per group each (n = 4–6). Each dot represents 1 mouse. *P < 0.05, **P < 0.01, Mann-Whitney U test with Bonferroni correction. Original magnification (H&E and IHC), ×10.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Activity Assay, Staining, MANN-WHITNEY

Colitis was induced in WT and Ptpn22–/– littermates by administration of 2.5% DSS for 7 days. (A) Colon specimens were analyzed for caspase-1, IL-1β, and IL-18 by Western blot. (B) NLRP3 was immunoprecipitated from whole colon specimens and analyzed for tyrosine phosphorylation and interaction with PTPN22. (C) Intestinal epithelial cell (IEC) fraction and lamina propria (LP) were analyzed for caspase-1, IL-1β, and IL-18 by Western blot. (D) Lamina propria cells and epithelial cells were analyzed for Ptpn22 mRNA levels normalized to Actb. (E) NLRP3 was precipitated from lamina propria or epithelial cells and analyzed for pTyr and PTPN22. Numbers below the Western blot images show results of densitometry, and each lane represents one mouse.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: Colitis was induced in WT and Ptpn22–/– littermates by administration of 2.5% DSS for 7 days. (A) Colon specimens were analyzed for caspase-1, IL-1β, and IL-18 by Western blot. (B) NLRP3 was immunoprecipitated from whole colon specimens and analyzed for tyrosine phosphorylation and interaction with PTPN22. (C) Intestinal epithelial cell (IEC) fraction and lamina propria (LP) were analyzed for caspase-1, IL-1β, and IL-18 by Western blot. (D) Lamina propria cells and epithelial cells were analyzed for Ptpn22 mRNA levels normalized to Actb. (E) NLRP3 was precipitated from lamina propria or epithelial cells and analyzed for pTyr and PTPN22. Numbers below the Western blot images show results of densitometry, and each lane represents one mouse.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Western Blot, Immunoprecipitation

Intestinal biopsies and serum samples from CD patients homozygous for the major (G) variant or heterozygous or homozygous for the minor (A) PTPN22 variant in SNP rs2476601 were analyzed for (A) PTPN22, NLRP3, or IL1B mRNA levels (normalized to ACTB); and (B) serum levels of IL-1β. Monocyte-derived dendritic cells from healthy controls (HC), CD patients homozygous for the G variant (GG), or CD patients heterozygous for the A variant (GA) were (C) primed for 16 hours with upLPS before activation with MSU for 6 hours prior to analysis for caspase-1 and IL-1β by Western blot; or (D) left untreated or primed for 16 hours with upLPS before analysis of PTPN22, NLRP3, or IL1B mRNA expression. Data are shown as values relative to nontreated controls and normalized to ACTB. Each dot/lane represents an individual patient; *P < 0.05, **P < 0.001 (Mann-Whitney U test with Bonferroni correction). Numbers below the Western blot images show results of densitometry.

Journal: The Journal of Clinical Investigation

Article Title: NLRP3 tyrosine phosphorylation is controlled by protein tyrosine phosphatase PTPN22

doi: 10.1172/JCI83669

Figure Lengend Snippet: Intestinal biopsies and serum samples from CD patients homozygous for the major (G) variant or heterozygous or homozygous for the minor (A) PTPN22 variant in SNP rs2476601 were analyzed for (A) PTPN22, NLRP3, or IL1B mRNA levels (normalized to ACTB); and (B) serum levels of IL-1β. Monocyte-derived dendritic cells from healthy controls (HC), CD patients homozygous for the G variant (GG), or CD patients heterozygous for the A variant (GA) were (C) primed for 16 hours with upLPS before activation with MSU for 6 hours prior to analysis for caspase-1 and IL-1β by Western blot; or (D) left untreated or primed for 16 hours with upLPS before analysis of PTPN22, NLRP3, or IL1B mRNA expression. Data are shown as values relative to nontreated controls and normalized to ACTB. Each dot/lane represents an individual patient; *P < 0.05, **P < 0.001 (Mann-Whitney U test with Bonferroni correction). Numbers below the Western blot images show results of densitometry.

Article Snippet: Antibodies used were anti–caspase-3 (Cell Signaling Technologies [CST]; catalog 9662); anti-NLRP3 (Enzo Life Sciences; for mouse: clone Cryo-1, catalog 804-880; for human: Nalpy-3a, 804-881); anti-PTPN22 (Santa Cruz Biotechnology Inc.; T-16, 48922); anti-ASC (CST; E1E3I, 13833); anti–caspase-1 (CST; D7F10, 3866); anti–IL-1β (CST; 3A6, 12242); AIM2 (Santa Cruz Biotechnology Inc.; D14; sc-137967); NLRC4 (CST; D5Y8E, 12421); PTPN2 (Merck Millipore; CF-41D, PH03L).

Techniques: Variant Assay, Derivative Assay, Activation Assay, Western Blot, Expressing, MANN-WHITNEY