Virus:Article Title: Potentiation of macrophage Piezo1 by atherogenic 7-ketocholesterol.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies F4/80 Monoclonal Antibody (BM8) Alexa FluorTM 488 ThermoFisher RRID: AB_10376287 CD45-APCCY7 Biolegend RRID: AB_3113064 CD64-BV786 BD Biosciences RRID: AB_2740644 FOLR2-APC Biolegend RRID: AB_2721303 Piezo-1 Polyclonal Antibody ProteinTech RRID: AB_2231460 Goat anti-Rabbit Alexa FluorTM 488 antibody ThermoFisher RRID: AB_2576217 Bacterial and virus strains JM109 competent cells Promega Cat#L200B Biological samples Bone marrow differentiated macrophages This paper N/A Chemicals, peptides, and recombinant proteins DMEM, high glucose Gibco Cat#41965-039 GibcoTM DPBS, without calcium and magnesium Fisher Scientific Cat#12559069 GibcoTM OptiMEM I Fisher Scientific Cat15392402 Fetal bovine serum (FBS) HyClone SV30143.03 New born calf serum Sigma-Aldrich Cat#N4637 Lipoprotein Deficient Serum This paper N/A Penicillin/Streptomycin Gibco Cat#15140-122 Glutamax Gibco Cat#35050-038 Poly-L-Lysine Sigma-Aldrich Cat#P4832 Trypsin-EDTA Gibco Cat#25300-054 NormocinTM Invivogen Cat#ant-nr-1 Zeocin Invivogen Cat#ant-zn-1 Collagen I Gibco Cat#A108301 jetPEI (Polyplus) Ozyme Cat#101-40N LipofectamineTM RNAiMAX Invitrogen Cat#13778-150 Recombinant Mouse M-CSF Protein Gibco Cat#PMC2044 Lipopolysaccharides from Escherichia coli O111:B4 Sigma-Aldrich Cat#L2630 IFN-gamma Protein, Mouse MedChemExpress Cat#HY-P7071 IL-4 Protein, Mouse MedChemExpress Cat#HY-P70644 IL-13 Protein, Mouse MedChemExpress Cat#HY-P70460 Low-Density Lipoprotein (LDL) Fisher Scientific Cat#10442132 Oxidized LDL (oxLDL) Fisher Scientific Cat#15643308 7-ketocholesterol (7-KC) Sigma-Aldrich Cat#700015P Methyl-b-cyclodextrin (MbCD) Sigma-Aldrich Cat#C4555 Cholesterol-Water Soluble Sigma-Aldrich Cat#C4951 Yoda1 Tocris Cat#5586 InvitrogenTM Trypan blue Fisher Scientific Cat#11584486 InvitrogenTM Random Primers Fisher Scientific Cat#10646313 GoTaq(R) G2 DNA Polymerase, 500u Promega Cat#M7845 MemGlowTM NR12 S Membrane Polarity Probe TebuBio Cat#MG08 Ergosta-5,7,9(11),22-tetraen-3b-ol (DHE) Sigma-Aldrich Cat#E2634 b-mercapthoethanol Sigma-Aldrich Cat#M3148 (Continued on next page) 14 Cell Reports 44, 115542, April 22, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Red blood cell lysis buffer Sigma-Aldrich Cat#R7757 cis-4,7,10,13,16,19-Docosahexaenoic acid (DHA) Sigma-Aldrich Cat#2534 Wheat germ agglutinin (WGA; Tetramethylrhodamine conjugate) ThermoFisher Cat#W849 BODIPY 493/503 Fisher Scientific Cat#11540326 InvitrogenTM HCS CellMaskTM Stains Fisher Scientific Cat#10615695 Potassium bromide Sigma-Aldrich Cat#746444 DAPI Sigma-Aldrich Cat#D9542 Hoechst Sigma-Aldrich Cat#33342 L-a-lyso-phosphatidylcholine (LPC) Avanti Polar Lipids Cat#830071 sphingomyelin (SM) Avanti Polar Lipids Cat#860062 L-a-phosphatidylcholine (PC) Avanti Polar Lipids Cat#840051 L-a-phosphatidylserine (PS) Avanti Polar Lipids Cat#840032 L-a-phosphatidylinositol (PI) Avanti Polar Lipids Cat#840042 Cardiolipin (CL) Avanti Polar Lipids Cat#840012 L-a-phosphatidylethanolamine (PE) Avanti Polar Lipids Cat#840026 L-a-phosphatidylglycerol (PG) Avanti Polar Lipids Cat#841138 Ceramide (Cer) Avanti Polar Lipids Cat#860051 Mono-Sulfo Galactosyl(b) Ceramide (d18:1/17:0) (MSGCer) Avanti Polar Lipids Cat#860572 Glucosyl(b) Ceramide (d18:1/17:0) (HexCer) Avanti Polar Lipids Cat#860569 C24 Lactosyl(b) Ceramide (d18:1/24:0) (Hex2Cer) Avanti Polar Lipids Cat#860577 5-cholestén-3b-ol (ST) Sigma Cat#C8667 Cholest-5-en-3b-yl heptadecanoate (STE) Avanti Polar Lipids Cat#700186 Cholesteryl formate (ChoF) Sigma Cat#S448532 Arachidonic acid (FFA) Sigma Cat#A3611 1-oleoyl-rac-glycerol (MG) Avanti Polar Lipids Cat#330724 1,3-di(cis-9-octadecenoyl)-glycerol (DG1.3) Sigma Cat#D3627 1,2-dipalmitoyl-sn-glycerol (DG1.2) Avanti Polar Lipids Cat#800816 1,2,3-tri-(9Z-octadecenoyl)-glycerol (TG) Sigma Cat#T7140 HPLC-grade chloroform (CHCl3) Fisher Scientific Cat#10050090 HPLC-grade methanol (MeOH) Sigma Cat#34860 HPLC-grade hexane Honeywell Riedel-de-HaënTM Cat#34859 Toluene Sigma Cat#244511 Ethyl acetate (EtAc) VWR Cat#23880.290 1-propanol Sigma Cat#402893 Butylhydroxytoluene (BHT) Sigma Cat#34750 Ammonium molybdate tetrahydrate Sigma Cat#A7302 Ascorbic acid Sigma Cat#33034 Copper (II) sulfate pentahydrate Sigma Cat#C8027 Sodium dihydrogen phosphate dihydrate VWR Chemicals Cat#28011.291 Perchloric acid 65–71% Carlo Erba Cat#409193 Nitric acid 65% VWR Chemicals Cat#20429.245 Sulfuric acid 95% VWR Chemicals Cat#20700.298 Phosphoric acid Sigma Cat#1.00565 Glass HPTLC Silica gel 60 plates Merck/Millipore Cat#1.05641.0001 Collagenase D Sigma Cat#011088882001 Liberase DH Research Grade Sigma Cat#5401054001 Fura-2 a.m. ThermoFisher Cat#F1221 Ionomycin ThermoFisher Cat#I24222 (Continued on next page) Cell Reports 44, 115542, April 22, 2025 15
Recombinant:Article Title: Potentiation of macrophage Piezo1 by atherogenic 7-ketocholesterol.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies F4/80 Monoclonal Antibody (BM8) Alexa FluorTM 488 ThermoFisher RRID: AB_10376287 CD45-APCCY7 Biolegend RRID: AB_3113064 CD64-BV786 BD Biosciences RRID: AB_2740644 FOLR2-APC Biolegend RRID: AB_2721303 Piezo-1 Polyclonal Antibody ProteinTech RRID: AB_2231460 Goat anti-Rabbit Alexa FluorTM 488 antibody ThermoFisher RRID: AB_2576217 Bacterial and virus strains JM109 competent cells Promega Cat#L200B Biological samples Bone marrow differentiated macrophages This paper N/A Chemicals, peptides, and recombinant proteins DMEM, high glucose Gibco Cat#41965-039 GibcoTM DPBS, without calcium and magnesium Fisher Scientific Cat#12559069 GibcoTM OptiMEM I Fisher Scientific Cat15392402 Fetal bovine serum (FBS) HyClone SV30143.03 New born calf serum Sigma-Aldrich Cat#N4637 Lipoprotein Deficient Serum This paper N/A Penicillin/Streptomycin Gibco Cat#15140-122 Glutamax Gibco Cat#35050-038 Poly-L-Lysine Sigma-Aldrich Cat#P4832 Trypsin-EDTA Gibco Cat#25300-054 NormocinTM Invivogen Cat#ant-nr-1 Zeocin Invivogen Cat#ant-zn-1 Collagen I Gibco Cat#A108301 jetPEI (Polyplus) Ozyme Cat#101-40N LipofectamineTM RNAiMAX Invitrogen Cat#13778-150 Recombinant Mouse M-CSF Protein Gibco Cat#PMC2044 Lipopolysaccharides from Escherichia coli O111:B4 Sigma-Aldrich Cat#L2630 IFN-gamma Protein, Mouse MedChemExpress Cat#HY-P7071 IL-4 Protein, Mouse MedChemExpress Cat#HY-P70644 IL-13 Protein, Mouse MedChemExpress Cat#HY-P70460 Low-Density Lipoprotein (LDL) Fisher Scientific Cat#10442132 Oxidized LDL (oxLDL) Fisher Scientific Cat#15643308 7-ketocholesterol (7-KC) Sigma-Aldrich Cat#700015P Methyl-b-cyclodextrin (MbCD) Sigma-Aldrich Cat#C4555 Cholesterol-Water Soluble Sigma-Aldrich Cat#C4951 Yoda1 Tocris Cat#5586 InvitrogenTM Trypan blue Fisher Scientific Cat#11584486 InvitrogenTM Random Primers Fisher Scientific Cat#10646313 GoTaq(R) G2 DNA Polymerase, 500u Promega Cat#M7845 MemGlowTM NR12 S Membrane Polarity Probe TebuBio Cat#MG08 Ergosta-5,7,9(11),22-tetraen-3b-ol (DHE) Sigma-Aldrich Cat#E2634 b-mercapthoethanol Sigma-Aldrich Cat#M3148 (Continued on next page) 14 Cell Reports 44, 115542, April 22, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Red blood cell lysis buffer Sigma-Aldrich Cat#R7757 cis-4,7,10,13,16,19-Docosahexaenoic acid (DHA) Sigma-Aldrich Cat#2534 Wheat germ agglutinin (WGA; Tetramethylrhodamine conjugate) ThermoFisher Cat#W849 BODIPY 493/503 Fisher Scientific Cat#11540326 InvitrogenTM HCS CellMaskTM Stains Fisher Scientific Cat#10615695 Potassium bromide Sigma-Aldrich Cat#746444 DAPI Sigma-Aldrich Cat#D9542 Hoechst Sigma-Aldrich Cat#33342 L-a-lyso-phosphatidylcholine (LPC) Avanti Polar Lipids Cat#830071 sphingomyelin (SM) Avanti Polar Lipids Cat#860062 L-a-phosphatidylcholine (PC) Avanti Polar Lipids Cat#840051 L-a-phosphatidylserine (PS) Avanti Polar Lipids Cat#840032 L-a-phosphatidylinositol (PI) Avanti Polar Lipids Cat#840042 Cardiolipin (CL) Avanti Polar Lipids Cat#840012 L-a-phosphatidylethanolamine (PE) Avanti Polar Lipids Cat#840026 L-a-phosphatidylglycerol (PG) Avanti Polar Lipids Cat#841138 Ceramide (Cer) Avanti Polar Lipids Cat#860051 Mono-Sulfo Galactosyl(b) Ceramide (d18:1/17:0) (MSGCer) Avanti Polar Lipids Cat#860572 Glucosyl(b) Ceramide (d18:1/17:0) (HexCer) Avanti Polar Lipids Cat#860569 C24 Lactosyl(b) Ceramide (d18:1/24:0) (Hex2Cer) Avanti Polar Lipids Cat#860577 5-cholestén-3b-ol (ST) Sigma Cat#C8667 Cholest-5-en-3b-yl heptadecanoate (STE) Avanti Polar Lipids Cat#700186 Cholesteryl formate (ChoF) Sigma Cat#S448532 Arachidonic acid (FFA) Sigma Cat#A3611 1-oleoyl-rac-glycerol (MG) Avanti Polar Lipids Cat#330724 1,3-di(cis-9-octadecenoyl)-glycerol (DG1.3) Sigma Cat#D3627 1,2-dipalmitoyl-sn-glycerol (DG1.2) Avanti Polar Lipids Cat#800816 1,2,3-tri-(9Z-octadecenoyl)-glycerol (TG) Sigma Cat#T7140 HPLC-grade chloroform (CHCl3) Fisher Scientific Cat#10050090 HPLC-grade methanol (MeOH) Sigma Cat#34860 HPLC-grade hexane Honeywell Riedel-de-HaënTM Cat#34859 Toluene Sigma Cat#244511 Ethyl acetate (EtAc) VWR Cat#23880.290 1-propanol Sigma Cat#402893 Butylhydroxytoluene (BHT) Sigma Cat#34750 Ammonium molybdate tetrahydrate Sigma Cat#A7302 Ascorbic acid Sigma Cat#33034 Copper (II) sulfate pentahydrate Sigma Cat#C8027 Sodium dihydrogen phosphate dihydrate VWR Chemicals Cat#28011.291 Perchloric acid 65–71% Carlo Erba Cat#409193 Nitric acid 65% VWR Chemicals Cat#20429.245 Sulfuric acid 95% VWR Chemicals Cat#20700.298 Phosphoric acid Sigma Cat#1.00565 Glass HPTLC Silica gel 60 plates Merck/Millipore Cat#1.05641.0001 Collagenase D Sigma Cat#011088882001 Liberase DH Research Grade Sigma Cat#5401054001 Fura-2 a.m. ThermoFisher Cat#F1221 Ionomycin ThermoFisher Cat#I24222 (Continued on next page) Cell Reports 44, 115542, April 22, 2025 15
Membrane:Article Title: Potentiation of macrophage Piezo1 by atherogenic 7-ketocholesterol.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies F4/80 Monoclonal Antibody (BM8) Alexa FluorTM 488 ThermoFisher RRID: AB_10376287 CD45-APCCY7 Biolegend RRID: AB_3113064 CD64-BV786 BD Biosciences RRID: AB_2740644 FOLR2-APC Biolegend RRID: AB_2721303 Piezo-1 Polyclonal Antibody ProteinTech RRID: AB_2231460 Goat anti-Rabbit Alexa FluorTM 488 antibody ThermoFisher RRID: AB_2576217 Bacterial and virus strains JM109 competent cells Promega Cat#L200B Biological samples Bone marrow differentiated macrophages This paper N/A Chemicals, peptides, and recombinant proteins DMEM, high glucose Gibco Cat#41965-039 GibcoTM DPBS, without calcium and magnesium Fisher Scientific Cat#12559069 GibcoTM OptiMEM I Fisher Scientific Cat15392402 Fetal bovine serum (FBS) HyClone SV30143.03 New born calf serum Sigma-Aldrich Cat#N4637 Lipoprotein Deficient Serum This paper N/A Penicillin/Streptomycin Gibco Cat#15140-122 Glutamax Gibco Cat#35050-038 Poly-L-Lysine Sigma-Aldrich Cat#P4832 Trypsin-EDTA Gibco Cat#25300-054 NormocinTM Invivogen Cat#ant-nr-1 Zeocin Invivogen Cat#ant-zn-1 Collagen I Gibco Cat#A108301 jetPEI (Polyplus) Ozyme Cat#101-40N LipofectamineTM RNAiMAX Invitrogen Cat#13778-150 Recombinant Mouse M-CSF Protein Gibco Cat#PMC2044 Lipopolysaccharides from Escherichia coli O111:B4 Sigma-Aldrich Cat#L2630 IFN-gamma Protein, Mouse MedChemExpress Cat#HY-P7071 IL-4 Protein, Mouse MedChemExpress Cat#HY-P70644 IL-13 Protein, Mouse MedChemExpress Cat#HY-P70460 Low-Density Lipoprotein (LDL) Fisher Scientific Cat#10442132 Oxidized LDL (oxLDL) Fisher Scientific Cat#15643308 7-ketocholesterol (7-KC) Sigma-Aldrich Cat#700015P Methyl-b-cyclodextrin (MbCD) Sigma-Aldrich Cat#C4555 Cholesterol-Water Soluble Sigma-Aldrich Cat#C4951 Yoda1 Tocris Cat#5586 InvitrogenTM Trypan blue Fisher Scientific Cat#11584486 InvitrogenTM Random Primers Fisher Scientific Cat#10646313 GoTaq(R) G2 DNA Polymerase, 500u Promega Cat#M7845 MemGlowTM NR12 S Membrane Polarity Probe TebuBio Cat#MG08 Ergosta-5,7,9(11),22-tetraen-3b-ol (DHE) Sigma-Aldrich Cat#E2634 b-mercapthoethanol Sigma-Aldrich Cat#M3148 (Continued on next page) 14 Cell Reports 44, 115542, April 22, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Red blood cell lysis buffer Sigma-Aldrich Cat#R7757 cis-4,7,10,13,16,19-Docosahexaenoic acid (DHA) Sigma-Aldrich Cat#2534 Wheat germ agglutinin (WGA; Tetramethylrhodamine conjugate) ThermoFisher Cat#W849 BODIPY 493/503 Fisher Scientific Cat#11540326 InvitrogenTM HCS CellMaskTM Stains Fisher Scientific Cat#10615695 Potassium bromide Sigma-Aldrich Cat#746444 DAPI Sigma-Aldrich Cat#D9542 Hoechst Sigma-Aldrich Cat#33342 L-a-lyso-phosphatidylcholine (LPC) Avanti Polar Lipids Cat#830071 sphingomyelin (SM) Avanti Polar Lipids Cat#860062 L-a-phosphatidylcholine (PC) Avanti Polar Lipids Cat#840051 L-a-phosphatidylserine (PS) Avanti Polar Lipids Cat#840032 L-a-phosphatidylinositol (PI) Avanti Polar Lipids Cat#840042 Cardiolipin (CL) Avanti Polar Lipids Cat#840012 L-a-phosphatidylethanolamine (PE) Avanti Polar Lipids Cat#840026 L-a-phosphatidylglycerol (PG) Avanti Polar Lipids Cat#841138 Ceramide (Cer) Avanti Polar Lipids Cat#860051 Mono-Sulfo Galactosyl(b) Ceramide (d18:1/17:0) (MSGCer) Avanti Polar Lipids Cat#860572 Glucosyl(b) Ceramide (d18:1/17:0) (HexCer) Avanti Polar Lipids Cat#860569 C24 Lactosyl(b) Ceramide (d18:1/24:0) (Hex2Cer) Avanti Polar Lipids Cat#860577 5-cholestén-3b-ol (ST) Sigma Cat#C8667 Cholest-5-en-3b-yl heptadecanoate (STE) Avanti Polar Lipids Cat#700186 Cholesteryl formate (ChoF) Sigma Cat#S448532 Arachidonic acid (FFA) Sigma Cat#A3611 1-oleoyl-rac-glycerol (MG) Avanti Polar Lipids Cat#330724 1,3-di(cis-9-octadecenoyl)-glycerol (DG1.3) Sigma Cat#D3627 1,2-dipalmitoyl-sn-glycerol (DG1.2) Avanti Polar Lipids Cat#800816 1,2,3-tri-(9Z-octadecenoyl)-glycerol (TG) Sigma Cat#T7140 HPLC-grade chloroform (CHCl3) Fisher Scientific Cat#10050090 HPLC-grade methanol (MeOH) Sigma Cat#34860 HPLC-grade hexane Honeywell Riedel-de-HaënTM Cat#34859 Toluene Sigma Cat#244511 Ethyl acetate (EtAc) VWR Cat#23880.290 1-propanol Sigma Cat#402893 Butylhydroxytoluene (BHT) Sigma Cat#34750 Ammonium molybdate tetrahydrate Sigma Cat#A7302 Ascorbic acid Sigma Cat#33034 Copper (II) sulfate pentahydrate Sigma Cat#C8027 Sodium dihydrogen phosphate dihydrate VWR Chemicals Cat#28011.291 Perchloric acid 65–71% Carlo Erba Cat#409193 Nitric acid 65% VWR Chemicals Cat#20429.245 Sulfuric acid 95% VWR Chemicals Cat#20700.298 Phosphoric acid Sigma Cat#1.00565 Glass HPTLC Silica gel 60 plates Merck/Millipore Cat#1.05641.0001 Collagenase D Sigma Cat#011088882001 Liberase DH Research Grade Sigma Cat#5401054001 Fura-2 a.m. ThermoFisher Cat#F1221 Ionomycin ThermoFisher Cat#I24222 (Continued on next page) Cell Reports 44, 115542, April 22, 2025 15
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