Cell Culture:Article Title: Core passive and facultative mTOR-mediated mechanisms coordinate mammalian protein synthesis and decay.
Article Snippet: Cells were passaged by trypsinization (Sigma, T4049-100ML) every 2-3 days and maintained at a confluency < 80%. .. E14 and CGR8 mouse embryonic stem cell lines were routinely cultured in GMEM (Sigma-Aldrich, G5154) supplemented with 10% ES cell-qualified fetal bovine serum (Gibco, 16141–079), 2 mM sodium pyruvate (Sigma-Aldrich, 113-24-6), 1% non-essential amino acids (Gibco, 11140–050), 1% penicillin/streptomycin (BioConcept, 4-01F00H), 2 mM L-glutamine (Gibco, 25030–024), 100 μM 2-mercaptoethanol (Sigma-Aldrich, 63689), leukaemia inhibitory factor (LIF — in-house), 3 μM GSK-3 Inhibitor XVI (Merck, 361559) and 0.8 μM PD184352 (Sigma-Aldrich, PZ0181), called hereafter 2i, at 37◦C, 5% CO2. .. Cells were plated on dishes coated with gelatin (Sigma-Aldrich, G9391).
Produced:Article Title: A reliable analytical procedure to determine the carbon isotopic signature of CO<sub>2</sub>-bearing COH fluids generated in petrological experiments
Article Snippet: .. The accuracy of the IRMS is checked periodically by analyzing the δ13C of CO2 produced by the digestion with phosphoric acid of calcium carbonate in-house standards (Merck® carbonate: δ13C =−41.20‰; MAQ Carrara marble: δ13C =+2.58‰). .. The δ13C of bulk OAA and Tetraselmis suecica, as well as of the residual char contained in the capsules of experiments COH210 and COH245, was determined by connecting the IRMS with a Thermo ScientificTM Flash 2000 organic elemental analyzer (OEA) set to nitrogen and carbon (NC) mode.
Calcium Carbonate:Article Title: A reliable analytical procedure to determine the carbon isotopic signature of CO<sub>2</sub>-bearing COH fluids generated in petrological experiments
Article Snippet: .. The accuracy of the IRMS is checked periodically by analyzing the δ13C of CO2 produced by the digestion with phosphoric acid of calcium carbonate in-house standards (Merck® carbonate: δ13C =−41.20‰; MAQ Carrara marble: δ13C =+2.58‰). .. The δ13C of bulk OAA and Tetraselmis suecica, as well as of the residual char contained in the capsules of experiments COH210 and COH245, was determined by connecting the IRMS with a Thermo ScientificTM Flash 2000 organic elemental analyzer (OEA) set to nitrogen and carbon (NC) mode.
Recombinant:Article Title: Core passive and facultative mTOR-mediated mechanisms coordinate mammalian protein synthesis and decay.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Ki67 BD Biosciences Cat# 550609; RRID: AB_393778 Anti-GFAP STEMCELL Technologies Cat# 60048; RRID: AB_3095092 Anti-MAP2 Sigma-Aldrich Cat# AB5622; RRID: AB_5622 Anti-TUBB eBioscience Cat# 14-4510; RRID: AB_2572876 Anti-SOX2 Cell Signaling Technology Cat# 23064S; RRID: AB_2714146 Anti-NANOG Thermo Fisher Scientific Cat# MA5-31459; RRID: AB_2787091 Anti-OCT4 Cell Signaling Technology Cat# 5677; RRID: AB_10547892 AlexaFluor647 secondary, anti-mouse Life Technologies Cat# A31571; RRID: AB_162542 AlexaFluor647 secondary, anti-rabbit Life Technologies Cat# A21443; RRID: AB_2535861 Chemicals, peptides, and recombinant proteins Dulbecco’s Modified Eagle Medium, high glucose, pyruvate Thermo Fisher Scientific Cat# 41966029 Fetal Bovine Serum, embryonic stem cell-qualified Thermo Fisher Scientific Cat# 16141079 Fetal Bovine Serum Thermo Fisher Scientific Cat# 10270106 Sodium pyruvate solution Sigma-Aldrich Cat# 113-24-6 Non-essential amino acids Gibco Cat# 11140-050 Penicillin/Streptomycin BioConcept Cat# 401F00H L-glutamine (2 mM) Gibco Cat# 25030-024 2-mercaptoethanol Sigma-Aldrich Cat# 63689 Leukaemia inhibitory factor (LIF) In-house N/A GSK-3 Inhibitor XVI Merck Cat# 361559 PD184352 Sigma-Aldrich Cat# PZ0181 Gelatin Sigma-Aldrich Cat# G9391 Trypsin Sigma-Aldrich Cat# T4049-100ML mTeSR Plus STEMCELL Technologies Cat# 100-0276 ReLeSR STEMCELL Technologies Cat# 05872 Corning Matrigel hESC-qualified matrix Corning Cat# 354277 Accutase Innovative Cell Technology Cat# AT104 BrainPhysTM Without Phenol-Red STEMCELL Technologies Cat# 05791 BrainPhysTM hPSC Neuron Kit STEMCELL Technologies Cat# 05795 STEMdiff Astrocyte Maturation Supplement A STEMCELL Technologies Cat# 100-0037 STEMdiff Astrocyte Maturation Supplement B STEMCELL Technologies Cat# 100-0017 Poly-L-Ornithine (PLO) Sigma-Aldrich Cat# P4957 Laminin Sigma-Aldrich Cat# L2020 ROCK inhibitor Y-27632 MilliporeSigma Cat# SCM075 Laminin 511 BioLamina Cat# LN511-0202 Fibronectin Sigma-Aldrich Cat# F4759 FluoroBriteTM DMEM Gibco Cat# A1896701 VECTASHIELD® HardSetTM Antifade Mounting Medium with DAPI Vector Laboratories Cat# H-1500-10 (Continued on next page) ll OPEN ACCESSArticle Cell Systems 17, 101456, January 21, 2026 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Critical commercial assays Proteasome-GloTM Chymotrypsin-Like Assay kit Promega Cat# G8621 RNeasy Mini kit QIAGEN Cat# 74104 PierceTM BCA Protein Assay Thermo Fisher Scientific Cat# 23225 Deposited data Label-free proteomics (LFQ) PRIDE Accession: PXD056601 dSILAC proteomics (NIH/3T3) PRIDE Accession: PXD061384 dSILAC proteomics (mESCs) PRIDE Accession: PXD061306 RNA-seq GEO Accession: GSE278929 Analysis code and data processing scripts GitHub https://github.com/UPSUTER/ProTuCo Experimental models: Cell lines NIH/3T3 Ueli Schibler, Geneva N/A
Article Title: Endothelial IL-36 receptor activation promotes vascular stability to limit pathological microvessel permeability in the CNS.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Polyclonal Anti-IL1Rrp2/IL-1R6 Novus Biologicals Cat#NBP1-81594; RRID:AB_11027148 Mouse Anti-Human VE-Cadherin (F-8) Santa Cruz Cat#sc-9989; RRID:AB_2077957 Rat Anti-Mouse CD31 (RM0032-1D12) Abcam Cat#ab56299; RRID:AB_940884 Rabbit Monoclonal anti-Vimentin (D21H3) Cell Signaling Cat#5741; RRID:AB_10695459 Isolectin GS-IB4 Alexa Fluor® 488 Conjugate Invitrogen Cat#I21411; RRID:AB_2314662 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) Abcam Cat#ab150160; RRID: AB_2756445 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 405) Abcam Cat# ab175652; RRID: AB_2687498 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) Invitrogen Cat# A-11032; RRID: AB_2534091 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) Abcam Cat# ab150077; RRID: AB_2630356 Rabbit monoclonal P-NFκB p65 (Ser536) Cell Signaling Cat#3033; RRID:AB_331284 Rabbit polyclonal Tricellulin (MARVELD2) Invitrogen Cat# 48–8400; RRID:AB_2533851 Rabbit monoclonal Claudin 5 Antibody Invitrogen Cat#MA5-32614; RRID: AB_2809891 Mouse Monoclonal β-actin antibody Sigma Aldrich Cat# A2228; RRID:AB_476697 Rabbit Monoclonal NF-κB p65 (D14E12) Cell Signaling Cat# 8242; RRID:AB_10859369 Bacterial and virus strains ClearColi® BL21(DE3) Lucigen/Biozym NC0487614 Biological samples Mouse: Primary Mouse Brain Microvascular Endothelial Cells (BMVECs) Isolated in-house N/A Chemicals, peptides, and recombinant proteins bisBenzimide H 33342 trihydrochloride (Hoechst) Sigma Aldrich Cat#B2261 Pierce TM 16% Formaldehyde (w/v), Methanol-free Fisher Scientific 28908 nickel-NTA agarose Expedeon ANN0010 PBS Merck D8662-500ML Imidazole Fisher Scientific 15805168 Coomassie Brilliant Blue Sigma Aldrich 1.15444 HEPES sodium salt Sigma Aldrich H3784 NaCl Sigma Aldrich 746398 DTT Merck 3860-OP Sodium Fluorescein Fisher Scientific 10714951 Goat serum Sigma Aldrich G9023 Triton TM X-100 Merck X100 IGEPAL® CA-630 Merck I8896 Sodium deoxycholate Merck 1.06504 CellTiter 96® AQueous MTS Reagent Powder Promega G1112 Recombinant human IL-36a 10 μg Peprotech 200-36a Recombinant human IL-36b 10 μg Peprotech 200-36b Protease Inhibitor Cocktail Merck P8340 (Continued on next page) Cell Reports 44, 116549, November 25, 2025 19 .. REAGENT or RESOURCE SOURCE IDENTIFIER Growth Factor Reduced Matrigel Corning 354230 Endothelial Cell Growth Medium MV2 PromoCell C-22022 Matrigel® Basement Membrane Matrix Corning 356234 Fluorescein isothiocyanate – Dextran 500000-Conjugate Sigma Aldrich 46947 Cycloheximide solution Sigma Aldrich C4859 Complete Classic Medium with Serum and CultureBoost TM Cell Systems 4Z0-500 Trypsin-EDTA 0.25% Gibco 25200056 Thrombin from bovine plasma Sigma-Aldrich T4648 Fibrinogen from bovine plasma Merck F8630-1G Fibronectin from human plasma (0.1% solution) Merck F0895-2MG Recombinant human VEGF Bio-Techne 293-VE-050/CF Tris hydrochloride Merck 10812846001 Sodium dodecyl sulfate Merck 436143 Bromophenol Blue Merck B0126 Critical commercial assays DuoSet® ELISA kits (VEGF, FGF1, FGF2) R&D Systems DY493-05, DY493-05, DY3139-05 In Situ Cell Death Detection Kit, TMR red Roche 12156792910 CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) Promega G5421 E.Z.N.A.® Total RNA Kit I Omega-BIO-TEK R6834-02 SensiFAST TM SYBR® No-ROX Kit Bioline/Meridian Bioscience BIO-98020 Deposited data Raw and analyzed data This paper GEO: GSE307365 Experimental models: Cell lines Human: Primary Retinal Microvascular Endothelial Cells (HRMECs) Cell-Systems ACBRI 181 Human: Primary Retinal Pericyte Cells Cell-Systems ACBRI-183 Human: Primary Retinal Endothelial Cells (HRECs) Innoprot P10880 Experimental models: Organisms/strains IL36 fl/fl mice Herná ndez-Santana et al. 53 N/A Tie2Cre mice The Jackson Laboratory JAX:016261 C57BL/6J The Jackson Laboratory JAX:000664 Il36rn− /− mice Gift from M. Kopf (ETH Zü rich) N/A Oligonucleotides For Primer Sequences for qPCR Analysis, See Method details N/A Software and algorithms ImageJ/Fiji NIH https://imagej.nih.gov/ij Imaris Oxford Instrument https://imaris.oxinst.com REAVER (Rapid Editable Analysis of Vessel Elements Routine) Corliss et al. 54 https://github.com/ktford/REAVER DESeq2 v1.38.3 Bioconductor https://bioconductor.org GSEA v4.3.3 Broad Institute https://www.gsea-msigdb.org/gsea/ index.jsp Hisat2 v2.05 Kim et al. 55 http://daehwankimlab.github.io/hisat2 StringTie v1.3.3b Pertea et al. 56 https://ccb.jhu.edu/software/stringtie (Continued on next page) 20 Cell Reports 44, 116549, November 25, 2025
Modification:Article Title: Core passive and facultative mTOR-mediated mechanisms coordinate mammalian protein synthesis and decay.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Ki67 BD Biosciences Cat# 550609; RRID: AB_393778 Anti-GFAP STEMCELL Technologies Cat# 60048; RRID: AB_3095092 Anti-MAP2 Sigma-Aldrich Cat# AB5622; RRID: AB_5622 Anti-TUBB eBioscience Cat# 14-4510; RRID: AB_2572876 Anti-SOX2 Cell Signaling Technology Cat# 23064S; RRID: AB_2714146 Anti-NANOG Thermo Fisher Scientific Cat# MA5-31459; RRID: AB_2787091 Anti-OCT4 Cell Signaling Technology Cat# 5677; RRID: AB_10547892 AlexaFluor647 secondary, anti-mouse Life Technologies Cat# A31571; RRID: AB_162542 AlexaFluor647 secondary, anti-rabbit Life Technologies Cat# A21443; RRID: AB_2535861 Chemicals, peptides, and recombinant proteins Dulbecco’s Modified Eagle Medium, high glucose, pyruvate Thermo Fisher Scientific Cat# 41966029 Fetal Bovine Serum, embryonic stem cell-qualified Thermo Fisher Scientific Cat# 16141079 Fetal Bovine Serum Thermo Fisher Scientific Cat# 10270106 Sodium pyruvate solution Sigma-Aldrich Cat# 113-24-6 Non-essential amino acids Gibco Cat# 11140-050 Penicillin/Streptomycin BioConcept Cat# 401F00H L-glutamine (2 mM) Gibco Cat# 25030-024 2-mercaptoethanol Sigma-Aldrich Cat# 63689 Leukaemia inhibitory factor (LIF) In-house N/A GSK-3 Inhibitor XVI Merck Cat# 361559 PD184352 Sigma-Aldrich Cat# PZ0181 Gelatin Sigma-Aldrich Cat# G9391 Trypsin Sigma-Aldrich Cat# T4049-100ML mTeSR Plus STEMCELL Technologies Cat# 100-0276 ReLeSR STEMCELL Technologies Cat# 05872 Corning Matrigel hESC-qualified matrix Corning Cat# 354277 Accutase Innovative Cell Technology Cat# AT104 BrainPhysTM Without Phenol-Red STEMCELL Technologies Cat# 05791 BrainPhysTM hPSC Neuron Kit STEMCELL Technologies Cat# 05795 STEMdiff Astrocyte Maturation Supplement A STEMCELL Technologies Cat# 100-0037 STEMdiff Astrocyte Maturation Supplement B STEMCELL Technologies Cat# 100-0017 Poly-L-Ornithine (PLO) Sigma-Aldrich Cat# P4957 Laminin Sigma-Aldrich Cat# L2020 ROCK inhibitor Y-27632 MilliporeSigma Cat# SCM075 Laminin 511 BioLamina Cat# LN511-0202 Fibronectin Sigma-Aldrich Cat# F4759 FluoroBriteTM DMEM Gibco Cat# A1896701 VECTASHIELD® HardSetTM Antifade Mounting Medium with DAPI Vector Laboratories Cat# H-1500-10 (Continued on next page) ll OPEN ACCESSArticle Cell Systems 17, 101456, January 21, 2026 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Critical commercial assays Proteasome-GloTM Chymotrypsin-Like Assay kit Promega Cat# G8621 RNeasy Mini kit QIAGEN Cat# 74104 PierceTM BCA Protein Assay Thermo Fisher Scientific Cat# 23225 Deposited data Label-free proteomics (LFQ) PRIDE Accession: PXD056601 dSILAC proteomics (NIH/3T3) PRIDE Accession: PXD061384 dSILAC proteomics (mESCs) PRIDE Accession: PXD061306 RNA-seq GEO Accession: GSE278929 Analysis code and data processing scripts GitHub https://github.com/UPSUTER/ProTuCo Experimental models: Cell lines NIH/3T3 Ueli Schibler, Geneva N/A
Derivative Assay:Article Title: Learning the All-Atom Equilibrium Distribution of Biomolecular Interactions at Scale
Article Snippet: .. To assess protein-ligand dynamics, we utilized three distinct test sets: a diverse held-out set of PDB systems to demonstrate broad chemical generalizability, the JACS & Merck [ ] datasets—industrial standards for structure-activity relationship (SAR) analysis—to capture subtle ligand-induced conformational changes and an in-house dataset derived from our internal drug discovery pipelines. ..
Drug discovery:Article Title: Learning the All-Atom Equilibrium Distribution of Biomolecular Interactions at Scale
Article Snippet: .. To assess protein-ligand dynamics, we utilized three distinct test sets: a diverse held-out set of PDB systems to demonstrate broad chemical generalizability, the JACS & Merck [ ] datasets—industrial standards for structure-activity relationship (SAR) analysis—to capture subtle ligand-induced conformational changes and an in-house dataset derived from our internal drug discovery pipelines. ..
Virus:Article Title: Endothelial IL-36 receptor activation promotes vascular stability to limit pathological microvessel permeability in the CNS.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Polyclonal Anti-IL1Rrp2/IL-1R6 Novus Biologicals Cat#NBP1-81594; RRID:AB_11027148 Mouse Anti-Human VE-Cadherin (F-8) Santa Cruz Cat#sc-9989; RRID:AB_2077957 Rat Anti-Mouse CD31 (RM0032-1D12) Abcam Cat#ab56299; RRID:AB_940884 Rabbit Monoclonal anti-Vimentin (D21H3) Cell Signaling Cat#5741; RRID:AB_10695459 Isolectin GS-IB4 Alexa Fluor® 488 Conjugate Invitrogen Cat#I21411; RRID:AB_2314662 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) Abcam Cat#ab150160; RRID: AB_2756445 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 405) Abcam Cat# ab175652; RRID: AB_2687498 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) Invitrogen Cat# A-11032; RRID: AB_2534091 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) Abcam Cat# ab150077; RRID: AB_2630356 Rabbit monoclonal P-NFκB p65 (Ser536) Cell Signaling Cat#3033; RRID:AB_331284 Rabbit polyclonal Tricellulin (MARVELD2) Invitrogen Cat# 48–8400; RRID:AB_2533851 Rabbit monoclonal Claudin 5 Antibody Invitrogen Cat#MA5-32614; RRID: AB_2809891 Mouse Monoclonal β-actin antibody Sigma Aldrich Cat# A2228; RRID:AB_476697 Rabbit Monoclonal NF-κB p65 (D14E12) Cell Signaling Cat# 8242; RRID:AB_10859369 Bacterial and virus strains ClearColi® BL21(DE3) Lucigen/Biozym NC0487614 Biological samples Mouse: Primary Mouse Brain Microvascular Endothelial Cells (BMVECs) Isolated in-house N/A Chemicals, peptides, and recombinant proteins bisBenzimide H 33342 trihydrochloride (Hoechst) Sigma Aldrich Cat#B2261 Pierce TM 16% Formaldehyde (w/v), Methanol-free Fisher Scientific 28908 nickel-NTA agarose Expedeon ANN0010 PBS Merck D8662-500ML Imidazole Fisher Scientific 15805168 Coomassie Brilliant Blue Sigma Aldrich 1.15444 HEPES sodium salt Sigma Aldrich H3784 NaCl Sigma Aldrich 746398 DTT Merck 3860-OP Sodium Fluorescein Fisher Scientific 10714951 Goat serum Sigma Aldrich G9023 Triton TM X-100 Merck X100 IGEPAL® CA-630 Merck I8896 Sodium deoxycholate Merck 1.06504 CellTiter 96® AQueous MTS Reagent Powder Promega G1112 Recombinant human IL-36a 10 μg Peprotech 200-36a Recombinant human IL-36b 10 μg Peprotech 200-36b Protease Inhibitor Cocktail Merck P8340 (Continued on next page) Cell Reports 44, 116549, November 25, 2025 19 .. REAGENT or RESOURCE SOURCE IDENTIFIER Growth Factor Reduced Matrigel Corning 354230 Endothelial Cell Growth Medium MV2 PromoCell C-22022 Matrigel® Basement Membrane Matrix Corning 356234 Fluorescein isothiocyanate – Dextran 500000-Conjugate Sigma Aldrich 46947 Cycloheximide solution Sigma Aldrich C4859 Complete Classic Medium with Serum and CultureBoost TM Cell Systems 4Z0-500 Trypsin-EDTA 0.25% Gibco 25200056 Thrombin from bovine plasma Sigma-Aldrich T4648 Fibrinogen from bovine plasma Merck F8630-1G Fibronectin from human plasma (0.1% solution) Merck F0895-2MG Recombinant human VEGF Bio-Techne 293-VE-050/CF Tris hydrochloride Merck 10812846001 Sodium dodecyl sulfate Merck 436143 Bromophenol Blue Merck B0126 Critical commercial assays DuoSet® ELISA kits (VEGF, FGF1, FGF2) R&D Systems DY493-05, DY493-05, DY3139-05 In Situ Cell Death Detection Kit, TMR red Roche 12156792910 CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) Promega G5421 E.Z.N.A.® Total RNA Kit I Omega-BIO-TEK R6834-02 SensiFAST TM SYBR® No-ROX Kit Bioline/Meridian Bioscience BIO-98020 Deposited data Raw and analyzed data This paper GEO: GSE307365 Experimental models: Cell lines Human: Primary Retinal Microvascular Endothelial Cells (HRMECs) Cell-Systems ACBRI 181 Human: Primary Retinal Pericyte Cells Cell-Systems ACBRI-183 Human: Primary Retinal Endothelial Cells (HRECs) Innoprot P10880 Experimental models: Organisms/strains IL36 fl/fl mice Herná ndez-Santana et al. 53 N/A Tie2Cre mice The Jackson Laboratory JAX:016261 C57BL/6J The Jackson Laboratory JAX:000664 Il36rn− /− mice Gift from M. Kopf (ETH Zü rich) N/A Oligonucleotides For Primer Sequences for qPCR Analysis, See Method details N/A Software and algorithms ImageJ/Fiji NIH https://imagej.nih.gov/ij Imaris Oxford Instrument https://imaris.oxinst.com REAVER (Rapid Editable Analysis of Vessel Elements Routine) Corliss et al. 54 https://github.com/ktford/REAVER DESeq2 v1.38.3 Bioconductor https://bioconductor.org GSEA v4.3.3 Broad Institute https://www.gsea-msigdb.org/gsea/ index.jsp Hisat2 v2.05 Kim et al. 55 http://daehwankimlab.github.io/hisat2 StringTie v1.3.3b Pertea et al. 56 https://ccb.jhu.edu/software/stringtie (Continued on next page) 20 Cell Reports 44, 116549, November 25, 2025
Isolation:Article Title: Endothelial IL-36 receptor activation promotes vascular stability to limit pathological microvessel permeability in the CNS.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Polyclonal Anti-IL1Rrp2/IL-1R6 Novus Biologicals Cat#NBP1-81594; RRID:AB_11027148 Mouse Anti-Human VE-Cadherin (F-8) Santa Cruz Cat#sc-9989; RRID:AB_2077957 Rat Anti-Mouse CD31 (RM0032-1D12) Abcam Cat#ab56299; RRID:AB_940884 Rabbit Monoclonal anti-Vimentin (D21H3) Cell Signaling Cat#5741; RRID:AB_10695459 Isolectin GS-IB4 Alexa Fluor® 488 Conjugate Invitrogen Cat#I21411; RRID:AB_2314662 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) Abcam Cat#ab150160; RRID: AB_2756445 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 405) Abcam Cat# ab175652; RRID: AB_2687498 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) Invitrogen Cat# A-11032; RRID: AB_2534091 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) Abcam Cat# ab150077; RRID: AB_2630356 Rabbit monoclonal P-NFκB p65 (Ser536) Cell Signaling Cat#3033; RRID:AB_331284 Rabbit polyclonal Tricellulin (MARVELD2) Invitrogen Cat# 48–8400; RRID:AB_2533851 Rabbit monoclonal Claudin 5 Antibody Invitrogen Cat#MA5-32614; RRID: AB_2809891 Mouse Monoclonal β-actin antibody Sigma Aldrich Cat# A2228; RRID:AB_476697 Rabbit Monoclonal NF-κB p65 (D14E12) Cell Signaling Cat# 8242; RRID:AB_10859369 Bacterial and virus strains ClearColi® BL21(DE3) Lucigen/Biozym NC0487614 Biological samples Mouse: Primary Mouse Brain Microvascular Endothelial Cells (BMVECs) Isolated in-house N/A Chemicals, peptides, and recombinant proteins bisBenzimide H 33342 trihydrochloride (Hoechst) Sigma Aldrich Cat#B2261 Pierce TM 16% Formaldehyde (w/v), Methanol-free Fisher Scientific 28908 nickel-NTA agarose Expedeon ANN0010 PBS Merck D8662-500ML Imidazole Fisher Scientific 15805168 Coomassie Brilliant Blue Sigma Aldrich 1.15444 HEPES sodium salt Sigma Aldrich H3784 NaCl Sigma Aldrich 746398 DTT Merck 3860-OP Sodium Fluorescein Fisher Scientific 10714951 Goat serum Sigma Aldrich G9023 Triton TM X-100 Merck X100 IGEPAL® CA-630 Merck I8896 Sodium deoxycholate Merck 1.06504 CellTiter 96® AQueous MTS Reagent Powder Promega G1112 Recombinant human IL-36a 10 μg Peprotech 200-36a Recombinant human IL-36b 10 μg Peprotech 200-36b Protease Inhibitor Cocktail Merck P8340 (Continued on next page) Cell Reports 44, 116549, November 25, 2025 19 .. REAGENT or RESOURCE SOURCE IDENTIFIER Growth Factor Reduced Matrigel Corning 354230 Endothelial Cell Growth Medium MV2 PromoCell C-22022 Matrigel® Basement Membrane Matrix Corning 356234 Fluorescein isothiocyanate – Dextran 500000-Conjugate Sigma Aldrich 46947 Cycloheximide solution Sigma Aldrich C4859 Complete Classic Medium with Serum and CultureBoost TM Cell Systems 4Z0-500 Trypsin-EDTA 0.25% Gibco 25200056 Thrombin from bovine plasma Sigma-Aldrich T4648 Fibrinogen from bovine plasma Merck F8630-1G Fibronectin from human plasma (0.1% solution) Merck F0895-2MG Recombinant human VEGF Bio-Techne 293-VE-050/CF Tris hydrochloride Merck 10812846001 Sodium dodecyl sulfate Merck 436143 Bromophenol Blue Merck B0126 Critical commercial assays DuoSet® ELISA kits (VEGF, FGF1, FGF2) R&D Systems DY493-05, DY493-05, DY3139-05 In Situ Cell Death Detection Kit, TMR red Roche 12156792910 CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) Promega G5421 E.Z.N.A.® Total RNA Kit I Omega-BIO-TEK R6834-02 SensiFAST TM SYBR® No-ROX Kit Bioline/Meridian Bioscience BIO-98020 Deposited data Raw and analyzed data This paper GEO: GSE307365 Experimental models: Cell lines Human: Primary Retinal Microvascular Endothelial Cells (HRMECs) Cell-Systems ACBRI 181 Human: Primary Retinal Pericyte Cells Cell-Systems ACBRI-183 Human: Primary Retinal Endothelial Cells (HRECs) Innoprot P10880 Experimental models: Organisms/strains IL36 fl/fl mice Herná ndez-Santana et al. 53 N/A Tie2Cre mice The Jackson Laboratory JAX:016261 C57BL/6J The Jackson Laboratory JAX:000664 Il36rn− /− mice Gift from M. Kopf (ETH Zü rich) N/A Oligonucleotides For Primer Sequences for qPCR Analysis, See Method details N/A Software and algorithms ImageJ/Fiji NIH https://imagej.nih.gov/ij Imaris Oxford Instrument https://imaris.oxinst.com REAVER (Rapid Editable Analysis of Vessel Elements Routine) Corliss et al. 54 https://github.com/ktford/REAVER DESeq2 v1.38.3 Bioconductor https://bioconductor.org GSEA v4.3.3 Broad Institute https://www.gsea-msigdb.org/gsea/ index.jsp Hisat2 v2.05 Kim et al. 55 http://daehwankimlab.github.io/hisat2 StringTie v1.3.3b Pertea et al. 56 https://ccb.jhu.edu/software/stringtie (Continued on next page) 20 Cell Reports 44, 116549, November 25, 2025
Protease Inhibitor:Article Title: Endothelial IL-36 receptor activation promotes vascular stability to limit pathological microvessel permeability in the CNS.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Polyclonal Anti-IL1Rrp2/IL-1R6 Novus Biologicals Cat#NBP1-81594; RRID:AB_11027148 Mouse Anti-Human VE-Cadherin (F-8) Santa Cruz Cat#sc-9989; RRID:AB_2077957 Rat Anti-Mouse CD31 (RM0032-1D12) Abcam Cat#ab56299; RRID:AB_940884 Rabbit Monoclonal anti-Vimentin (D21H3) Cell Signaling Cat#5741; RRID:AB_10695459 Isolectin GS-IB4 Alexa Fluor® 488 Conjugate Invitrogen Cat#I21411; RRID:AB_2314662 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) Abcam Cat#ab150160; RRID: AB_2756445 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 405) Abcam Cat# ab175652; RRID: AB_2687498 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) Invitrogen Cat# A-11032; RRID: AB_2534091 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) Abcam Cat# ab150077; RRID: AB_2630356 Rabbit monoclonal P-NFκB p65 (Ser536) Cell Signaling Cat#3033; RRID:AB_331284 Rabbit polyclonal Tricellulin (MARVELD2) Invitrogen Cat# 48–8400; RRID:AB_2533851 Rabbit monoclonal Claudin 5 Antibody Invitrogen Cat#MA5-32614; RRID: AB_2809891 Mouse Monoclonal β-actin antibody Sigma Aldrich Cat# A2228; RRID:AB_476697 Rabbit Monoclonal NF-κB p65 (D14E12) Cell Signaling Cat# 8242; RRID:AB_10859369 Bacterial and virus strains ClearColi® BL21(DE3) Lucigen/Biozym NC0487614 Biological samples Mouse: Primary Mouse Brain Microvascular Endothelial Cells (BMVECs) Isolated in-house N/A Chemicals, peptides, and recombinant proteins bisBenzimide H 33342 trihydrochloride (Hoechst) Sigma Aldrich Cat#B2261 Pierce TM 16% Formaldehyde (w/v), Methanol-free Fisher Scientific 28908 nickel-NTA agarose Expedeon ANN0010 PBS Merck D8662-500ML Imidazole Fisher Scientific 15805168 Coomassie Brilliant Blue Sigma Aldrich 1.15444 HEPES sodium salt Sigma Aldrich H3784 NaCl Sigma Aldrich 746398 DTT Merck 3860-OP Sodium Fluorescein Fisher Scientific 10714951 Goat serum Sigma Aldrich G9023 Triton TM X-100 Merck X100 IGEPAL® CA-630 Merck I8896 Sodium deoxycholate Merck 1.06504 CellTiter 96® AQueous MTS Reagent Powder Promega G1112 Recombinant human IL-36a 10 μg Peprotech 200-36a Recombinant human IL-36b 10 μg Peprotech 200-36b Protease Inhibitor Cocktail Merck P8340 (Continued on next page) Cell Reports 44, 116549, November 25, 2025 19 .. REAGENT or RESOURCE SOURCE IDENTIFIER Growth Factor Reduced Matrigel Corning 354230 Endothelial Cell Growth Medium MV2 PromoCell C-22022 Matrigel® Basement Membrane Matrix Corning 356234 Fluorescein isothiocyanate – Dextran 500000-Conjugate Sigma Aldrich 46947 Cycloheximide solution Sigma Aldrich C4859 Complete Classic Medium with Serum and CultureBoost TM Cell Systems 4Z0-500 Trypsin-EDTA 0.25% Gibco 25200056 Thrombin from bovine plasma Sigma-Aldrich T4648 Fibrinogen from bovine plasma Merck F8630-1G Fibronectin from human plasma (0.1% solution) Merck F0895-2MG Recombinant human VEGF Bio-Techne 293-VE-050/CF Tris hydrochloride Merck 10812846001 Sodium dodecyl sulfate Merck 436143 Bromophenol Blue Merck B0126 Critical commercial assays DuoSet® ELISA kits (VEGF, FGF1, FGF2) R&D Systems DY493-05, DY493-05, DY3139-05 In Situ Cell Death Detection Kit, TMR red Roche 12156792910 CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) Promega G5421 E.Z.N.A.® Total RNA Kit I Omega-BIO-TEK R6834-02 SensiFAST TM SYBR® No-ROX Kit Bioline/Meridian Bioscience BIO-98020 Deposited data Raw and analyzed data This paper GEO: GSE307365 Experimental models: Cell lines Human: Primary Retinal Microvascular Endothelial Cells (HRMECs) Cell-Systems ACBRI 181 Human: Primary Retinal Pericyte Cells Cell-Systems ACBRI-183 Human: Primary Retinal Endothelial Cells (HRECs) Innoprot P10880 Experimental models: Organisms/strains IL36 fl/fl mice Herná ndez-Santana et al. 53 N/A Tie2Cre mice The Jackson Laboratory JAX:016261 C57BL/6J The Jackson Laboratory JAX:000664 Il36rn− /− mice Gift from M. Kopf (ETH Zü rich) N/A Oligonucleotides For Primer Sequences for qPCR Analysis, See Method details N/A Software and algorithms ImageJ/Fiji NIH https://imagej.nih.gov/ij Imaris Oxford Instrument https://imaris.oxinst.com REAVER (Rapid Editable Analysis of Vessel Elements Routine) Corliss et al. 54 https://github.com/ktford/REAVER DESeq2 v1.38.3 Bioconductor https://bioconductor.org GSEA v4.3.3 Broad Institute https://www.gsea-msigdb.org/gsea/ index.jsp Hisat2 v2.05 Kim et al. 55 http://daehwankimlab.github.io/hisat2 StringTie v1.3.3b Pertea et al. 56 https://ccb.jhu.edu/software/stringtie (Continued on next page) 20 Cell Reports 44, 116549, November 25, 2025
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