Transfection:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Flow Cytometry:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Staining:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Electron Microscopy:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Lysis:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Isolation:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Single Cell:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Gene Expression:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
Reverse Transcription:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
BIA-KA:Article Title: A shear stress-responsive pathway in monocytes drives cardiopulmonary bypass-induced inflammation via spectrin/RAF1/store-operated calcium entry.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals and peptides RPMI-1640 Gibco Cat# 11875093 Fetal Bovine Serum, heat inactivated Gibco Cat# 16140-071 Penicillin/Streptomycin Gibco Cat# 15140122 DMEM Gibco Cat# 10566016 PBS Gibco Cat# 10010023 TransIT-Lenti Transfection Reagent Mirus Bio Cat# MIR 6603 FR180204 (ERK Inhibitor II) Cayman Cat# 15544 DMSO Alfa Aesar Cat# 42780 eBioscience TM Protein Transport Inhibitor Cocktail Thermo Fisher Cat# 00-4980-93 IC Fixation Buffer Thermo Fisher Cat# 00-8222-49 Permeabilization Buffer Thermo Fisher Cat# 00-8333-56 eBioscience TM Flow Cytometry Staining Buffer Thermo Fisher Cat# 00-4222-26 Proteinase K Zymo Cat# D3001-2-5 Polybrene MilliporeSigma Cat# TR-1003 Puromycin MedChemExpress Cat# HY-B1743 Doxycycline Thermo Fisher Cat# J60579.22 Propidium iodide MilliporeSigma Cat# P4170 Paraformaldehyde, methanol-free, 16% Electron Microscopy Sciences Cat# 15710 Triton X-100 (DNAse, RNAse, protease free) Acros Organics Cat# 327371000 RIPA lysis buffer Thermo Fisher Cat# 89900 SRC-selective tyrosine kinase inhibitor PP1 MilliporeSigma Cat# 5678091MG Ammonium Bicarbonate Thermo Fisher Cat# 393210010 Urea Thermo Fisher Cat# 29700 TCEP Thermo Fisher Cat# PG82080 Chloro-acetamide Thermo Fisher Cat# A15238.30 Trypsin/EDTA Gibco Cat# 25300054 Hydroxylamine Thermo Fisher Cat# A15398.0I Acetonitrile Thermo Fisher Cat# 047138.M1 Formic acid Thermo Fisher Cat# 28905 Sorafenib (Bay 43–9006) MedChemExpress Cat# HY-10201 Ulixertinib (BVD-523) MedChemExpress Cat# HY-15816 Zegocractin (CM-4620) MedChemExpress Cat# HY-101942 Critical commercial assays EasySep TM Direct Human PBMC Isolation Kit STEMCELL Technologies Cat# 19654 Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle 10x genomics Cat# PN-1000283 Chromium Fixed RNA Kit, Human Transcriptome 10x genomics Cat# PN-1000476 Dead cell removal kit Miltenyi Biotec Cat# 130-090-101 Quick-DNA/RNA Miniprep Kit Zymo Cat# D7005T Dead cell apoptosis kit, Alexa 488 w/Annexin V PI Invitrogen Cat# V13241 RNeasy® Mini Kit Qiagen Cat# 74104 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Nextera Library Prep Kit Illumina Cat# FC-131-1096 (Continued on next page) Cell Reports 45, 116903, February 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MinElute PCR purification Qiagen Cat# 28004 KAPA Library Quantification Kit KAPA Biosystems Cat# KK4824 CUTANA TM CUT&RUN Library Prep Kit EpiCypher Cat# 14-1001 Qubit 1X dsDNA HS Assay Kit Thermo Fisher Cat# Q33230 Pierce TM bicinchoninic acid (BCA) assay kit Thermo Fisher Cat# 23225 Duolink® In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Oligonucleotides qPCR primers for IL8, GAPDH, and RN18S1: see Table S2 This study N/A gRNA sequences for STIM1, ORAI1, SPTAN1, and RAF1 knockouts: see Table S3 This study N/A Plasmid lentiCRISPRv2 Addgene Cat# 73179-LV Lenti-iCas9-neo Addgene Cat# 85400 Lenti-multi-Guide plasmid Addgene Cat# 85401 pCDH-CMV-Nluc Addgene Cat#73038 pLenti CMV GFP Puro Addgene Cat# 658-5 pcDNA3-Venus-173-N-ORAI1 Addgene Cat# 87618 pcDNA3.1-STIM1-Venus-173-C Addgene Cat# 87619 pCDH-CMV-Nluc Addgene Cat# 73038 Biological samples Human peripheral blood mononuclear cells (PBMCs) This study N/A THP-1 cells ATCC TIB-202 Lenti-X 293T Takara Bio Cat# 632180 Human CD14 + primary monocytes Bloodworks Bio 4570–61 Antibodies IL8 antibody Invitrogen Cat# 12808842; RRID: AB_2784632 Anti-Ras antibody Abcam Cat# AB108602; RRID: AB_10891004 Raf1 antibody Abcam Cat# AB137435; RRID: AB_3674177 Goat anti-Mouse Alexa Fluor 488 Life Technologies Cat# A32723; RRID: AB_2633275 Goat anti-Rabbit, Alexa Fluor 594 Life Technologies Cat# A32740; RRID: AB_2762824 Anti-Spectrin alpha chain (nonerythroid) Antibody MilliporeSigma Cat# MAB1622; RRID: AB_94295 p-ERK1/2 antibody Cell Signaling Cat# 9101; RRID: AB_331646 ERK1/2 antibody Cell Signaling Cat# 4695; RRID: AB_390779 IRDye 680RD Goat anti-Mouse Li-Cor Cat# 926-68070; RRID: AB_10956588 IRDye 800CW Goat anti-Rabbit Li-Cor Cat# 926-32211; RRID: AB_621843 c-Jun Antibody Thermo Fisher Cat# MA5-15881; RRID: AB_11153582 β-actin Santa Cruz Cat# SC-47778; RRID: AB_626632 Deposited data snRNA-seq/snATAC-seq, ATAC, bulk-RNA, CUT&RUN data This paper NCBI GEO: GSE262146 CRISPRi screen data: See Data S1 This paper NCBI GEO: GSE213184 scRNA-seq data of drug treatment This paper NCBI GEO: GSE291575 Phospho-proteomics: See Data S2 This paper N/A Software and algorithms R 4.2.3 CRAN project https://www.r-project.org/ Seurat R package Version: 4.3.0.1 (Continued on next page) 16 Cell Reports 45, 116903, February 24, 2026
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