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    Bio-Rad protein g magnetic beads bio rad
    Protein G Magnetic Beads Bio Rad, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 640 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protein+g+magnetic+beads+bio+rad/SureBeads+Protein+G+Magnetic+Beads/pmc12376690-389-2-6
    Average 99 stars, based on 640 article reviews
    protein g magnetic beads bio rad - by Bioz Stars, 2026-09
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    Incubation:

    Article Title: Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex.
    Article Snippet: .. A total of 500 μg protein was incubated with vinculin antibody overnight at 4°C with rotation, and then 50 μL protein G magnetic beads (Bio‐Rad) was added. ..

    Article Title: Dual roles of HSP70 chaperone HSPA1 in quality control of nascent and newly synthesized proteins
    Article Snippet: To label and release nascent peptides by puromycin, 1 × 10 7 cells were collected and washed with cold PBS, followed by lysis in 500 μl of hypotonic buffer (supplemented with 20 μM MG132, 50 μM NEM, 1 mM DTT, 1× Pierce™ protease inhibitor cocktail and 2 mM ATP) on ice for 1 h with vortexing for 10 s every 10 min. Lysates were clarified by centrifugation at 9,400 g for 1 min. 100 μg/ml puromycin was added to the lysate, followed by incubation at room temperature for 1 h. 100 μl of the supernatant were removed (total lysate). .. To pull down puromycylated proteins, anti‐puromycin antibody (1:100) was added and incubated at 4°C for 4 h. 50 μl protein‐G magnetic beads (BIO‐RAD) blocked with 5% BSA and washed in lysis buffer twice were added, followed by overnight incubation. ..

    Article Title: Akkermansia Muciniphila Alleviates Severe Acute Pancreatitis via Amuc1409‐Ube2k‐Foxp3 Axis in Regulatory T Cells
    Article Snippet: .. 50 μL protein G magnetic beads (Bio‐Rad) prewashed with 0.1% PBST were added and incubated for another 4 h at 4 °C. ..

    Article Title: Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex
    Article Snippet: .. A total of 500 μg protein was incubated with vinculin antibody overnight at 4°C with rotation, and then 50 μL protein G magnetic beads (Bio‐Rad) was added. ..

    Magnetic Beads:

    Article Title: Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex.
    Article Snippet: .. A total of 500 μg protein was incubated with vinculin antibody overnight at 4°C with rotation, and then 50 μL protein G magnetic beads (Bio‐Rad) was added. ..

    Article Title: Dual roles of HSP70 chaperone HSPA1 in quality control of nascent and newly synthesized proteins
    Article Snippet: To label and release nascent peptides by puromycin, 1 × 10 7 cells were collected and washed with cold PBS, followed by lysis in 500 μl of hypotonic buffer (supplemented with 20 μM MG132, 50 μM NEM, 1 mM DTT, 1× Pierce™ protease inhibitor cocktail and 2 mM ATP) on ice for 1 h with vortexing for 10 s every 10 min. Lysates were clarified by centrifugation at 9,400 g for 1 min. 100 μg/ml puromycin was added to the lysate, followed by incubation at room temperature for 1 h. 100 μl of the supernatant were removed (total lysate). .. To pull down puromycylated proteins, anti‐puromycin antibody (1:100) was added and incubated at 4°C for 4 h. 50 μl protein‐G magnetic beads (BIO‐RAD) blocked with 5% BSA and washed in lysis buffer twice were added, followed by overnight incubation. ..

    Article Title: Akkermansia Muciniphila Alleviates Severe Acute Pancreatitis via Amuc1409‐Ube2k‐Foxp3 Axis in Regulatory T Cells
    Article Snippet: .. 50 μL protein G magnetic beads (Bio‐Rad) prewashed with 0.1% PBST were added and incubated for another 4 h at 4 °C. ..

    Article Title: CCR 5 deficiency impairs CD 4 + T‐cell memory responses and antigenic sensitivity through increased ceramide synthesis
    Article Snippet: .. Immune complexes were captured using Protein G Magnetic Beads (Bio‐Rad) and, after washing, eluted with 100 mM NaHCO 3 , 1% SDS; protein/DNA bonds were disrupted with proteinase K (10 μg/μl, 2 h, 62°C). ..

    Article Title: Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex
    Article Snippet: .. A total of 500 μg protein was incubated with vinculin antibody overnight at 4°C with rotation, and then 50 μL protein G magnetic beads (Bio‐Rad) was added. ..

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Article Title: Modulation of I Ks channel–PIP 2 interaction by PRMT1 plays a critical role in the control of cardiac repolarization
    Article Snippet: Upon clearance by centrifugation at 15,000 rpm for 15 min at 4°C, the lysates were incubated with anti‐Flag antibody (Sigma‐Aldrich) with gentle rotation overnight at 4°C. .. The immune complexes were then supplemented with protein G magnetic beads (Bio‐Rad) for 2 h at 4°C. ..

    Lysis:

    Article Title: Dual roles of HSP70 chaperone HSPA1 in quality control of nascent and newly synthesized proteins
    Article Snippet: To label and release nascent peptides by puromycin, 1 × 10 7 cells were collected and washed with cold PBS, followed by lysis in 500 μl of hypotonic buffer (supplemented with 20 μM MG132, 50 μM NEM, 1 mM DTT, 1× Pierce™ protease inhibitor cocktail and 2 mM ATP) on ice for 1 h with vortexing for 10 s every 10 min. Lysates were clarified by centrifugation at 9,400 g for 1 min. 100 μg/ml puromycin was added to the lysate, followed by incubation at room temperature for 1 h. 100 μl of the supernatant were removed (total lysate). .. To pull down puromycylated proteins, anti‐puromycin antibody (1:100) was added and incubated at 4°C for 4 h. 50 μl protein‐G magnetic beads (BIO‐RAD) blocked with 5% BSA and washed in lysis buffer twice were added, followed by overnight incubation. ..

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Transfection:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    RNA Extraction:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    cDNA Synthesis:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    SYBR Green Assay:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Selection:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Protease Inhibitor:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Bicinchoninic Acid Protein Assay:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2

    Membrane:

    Article Title: Stk24 protects against obesity-associated metabolic disorders by disrupting the NLRP3 inflammasome.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NLRP3 Adipogen International Cat# AG-20B-0014- C100 Mouse monoclonal anti-caspase-1 p20 Adipogen International Cat# AG-20B-0042- C100 Rabbit Polyclonal anti-ASC Adipogen International Cat# AG-20B-0006- C100 Rabbit monoclonal anti-p-ERK Cell Signaling Cat# 4370S;RRID:AB_2315112 Rabbit monoclonal anti-p-p38 Cell Signaling Cat# 9216; RRID:AB_331296 Rabbit monoclonal anti-p-p65 Cell Signaling Cat# 3033S; RRID:AB_331284 Rabbit monoclonal anti-p-JNK Cell Signaling Cat# 4668S; RRID:AB_823558 Rabbit monoclonal anti-p-PI3K Cell Signaling Cat# 4257S; RRID:AB_659889 PE anti-mouse F4/80 Antibody BioLegend Cat# 123110; RRID:AB_893486 Pacific Blue anti-mouse/human CD11b Antibody BioLegend Cat# 101224; RRID:AB_755986 FITC anti-mouse CD45.1 Antibody BioLegend Cat# 110706; RRID:AB_313495 APC anti-mouse CD45.2 Antibody BioLegend Cat# 109814; RRID:AB_389211 Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody BioLegend Cat# 108421; RRID:AB_493278 Rabbit monoclonal anti-H3K4me3 Millipore Cat# 07473; RRID:AB_1977252 Rabbit Polyclonal anti-H3K27me3 Millipore Cat# 07449; RRID:AB_310624 Rabbit monoclonal anti-Stk24 Abcam Cat# ab51137; RRID:AB_881248 Rabbit monoclonal anti-H3K9Ac Abcam Cat# ab4441; RRID:AB_2118292 Rabbit monoclonal anti-H3K27Ac Abcam Cat# ab4729; RRID:AB_2118291 Rabbit Polyclonal anti-GAPDH Diagbio Cat# db106 Mouse monoclonal anti-GFP Abclonal Cat# AE012; RRID:AB_2770402 Rabbit monoclonal anti-Flag Abclonal Cat# AE063; RRID:AB_2771920 Rabbit Polyclonal anti-Myc HUABIO Cat# R1208-1 Rabbit Polyclonal anti-HA HUABIO Cat# 0906-1 Biological Samples N/A N/A The peripheral blood samples The Second Affiliated Hospital, Zhejiang University College of Medicine N/A The omental adipose tissue and liver samples The China-Japan Friendship Hospital N/A Chemicals, peptides, and recombinant proteins LPS Sigma-Aldrich Cat# L2630 anti-flag mouse magnetic (M2) beads Sigma-Aldrich Cat# F1804 ATP InvivoGen Cat# tlrl-atpl nigericin InvivoGen Cat# tlrl-nig Imject Alum Thermo Scientific Cat# 77161 Lipofectamine 3000 Invitrogen Cat# L3000015 M-CSF Sigma-Aldrich Cat# H6916 Mouse recombinant IL-1b novoprotein Cat# C042 Mouse recombinant IL-4 novoprotein Cat# CK15 Human recombinant IL-1b novoprotein Cat# CG93 Mouse recombinant TNFa PeproTech Cat# 315-01A Human recombinant TNFa PeproTech Cat# 300-01A BAY11-7082 Beyotime Cat# SF0011 U0126 Beyotime Cat# S1901 (Continued on next page) e1 Cell Reports 35, 109161, May 25, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Rapamycin TargetMol Cat# T1537 INTERFERin@ Polyplus Transfection Cat# 409-10 CY-09 Dr. Rongbin Zhou N/A PEI Polyscience Cat# 19850 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778150 TRIeasyTM LS Total RNA Extraction Reagent Yeasen Cat# 19201ES60 Hifair II 1st Strand cDNA Synthesis Kit Yeasen Cat# 11119ES60 SYBR Green mix Yeasen Cat# 11199ES03 Collagenase II Sigma-Aldrich Cat# C6885 Human CD14 Positive Selection Kit STEMCELL Technologies Cat# 14758 Cell lysis buffer Cell Signaling Cat# 9803 Protease Inhibitor Sigma-Aldrich Cat# P2714 Phosphatase Inhibitor Sigma-Aldrich Cat# P2850 BCA protein assay kit Thermo Scientific Cat# 23250 Nitrocellulose membrane Bio-Rad Cat# 1703202 Protein G magnetic beads Bio-Rad Cat# 1614201 Tissue-Tek O.C.T. .. Compound Sakura Finetek Cat# 4583 Low-fat diet Research Diet Cat# D12450B High-fat diet Research Diet Cat# D12492 Critical commercial assays Mouse TNF-a ELISA Kit Invitrogen Cat# 88-7324-76 Mouse IL-6 ELISA Kit Invitrogen Cat# 88-7064-22 Mouse IL-1b ELISA Kit ebioscience Cat# 13-7112-68A Human IL-1b ELISA Kit ebioscience Cat# 33-7110-68 Deposited data The RNA sequencing data This paper NCBI Sequence Read Archive (PRJNA558987) Experimental models: cell lines 293T ATCC Cat# ACS-4500 THP-1 ATCC Cat# TIB-202 Experimental models: organisms/strains Stk24h/+ mice Dr. Dianqing Wu N/A CD45.1 (B6.SJL) Jackson Laboratory Cat# 002014 Oligonucleotides See Table S1 This paper N/A Recombinant DNA pCDNA3.1-Stk24–Flag This paper N/A pCDNA3.1-Stk24–K53R-Flag This paper N/A pCDNA3.1-Stk24–Myc This paper N/A pCDNA3.1-Stk24–1-277-Myc This paper N/A pCDNA3.1-Stk24–1-313-Myc This paper N/A pCDNA3.1-Stk24–C-Myc This paper N/A pCDNA3-NLRP3-Flag Dr. Rongbin Zhou N/A pCDNA3-ASC-Flag Dr. Rongbin Zhou N/A pCDNA3.1-NLRP3–PYD-Flag This paper N/A pCDNA3.1-NLRP3–NACHT-Flag This paper N/A pCDNA3.1-NLRP3–LRR-Flag This paper N/A (Continued on next page) Cell Reports 35, 109161, May 25, 2021 e2



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