Transfection:Article Title: Phenotypic pleiotropy of missense variants in human B cell confinement receptor P2RY8.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human TruStain FcX block Biolegend Cat# 422302; RRID:AB_2818986 InVivoPlus anti-mouse CD16/CD32 (mouse Fc block) Bio X Cell Cat# BE0307; RRID:AB_2736987 Armenian hamster anti-mouse CD3ε biotin (clone 145-2C11) Biolegend Cat# 100303, RRID:AB_312668 Syrian hamster anti-mouse anti-CD3ε AF700 (clone 500A2) Biolegend Cat# 152316; RRID:AB_2632713 Rat anti-mouse CD4 BV711 (clone GK1.5) Biolegend Cat# 100447; RRID:AB_2564586 Rat anti-mouse CD8a BV510 (clone 53-6.7) Biolegend Cat# 100752; RRID:AB_2563057 Rat anti-mouse CD11b BV570 (clone M1/70) Biolegend Cat# 101233; RRID:AB_10896949 Rat anti-mouse CD35 (CR1) biotin (clone 8C12) Fisher Scientific Cat# 553816; RRID:AB_2643602 Mouse anti-mouse CD45.1 APC (clone A20) Cytek Biosciences Cat# 20–0453; RRID:AB_2621575 Mouse anti-mouse CD45.1 PE (clone 104) Biolegend Cat# 109808; RRID:AB_313445 Rat anti-mouse/human CD45R/B220 BV785 (clone RA3-6B2) Biolegend Cat# 103246; RRID:AB_2563256 Hamster anti-mouse CD95 PE-Cy7 (clone Jo2) BD Biosciences Cat# 557653; RRID:AB_396768 Rat anti-mouse CD180 (clone RP/14) BD Biosciences Cat# 562191; RRID:AB_11153851 Rabbit anti-GFP AF488 (polyclonal) Thermo Fisher Scientific Cat# A21311; RRID:AB_221477 Rat anti-GFP AF488 (clone FM2-64G) Biolegend Cat # 338007; RRID:AB_2563287 Rat anti-mouse/human GL7 antigen Pacific Blue (clone GL7) Biolegend Cat# 144614; RRID:AB_2563292 Rat anti-mouse IgD AF647 (clone 11-26c.2a) Biolegend Cat# 405708; RRID:AB_893528 Rat anti-mouse IgD PerCP-Cy5.5 (clone 11-26c.2a) Biolegend Cat# 405710; RRID:AB_1575113 Rat anti-OX56 antigen biotin Contract production Bio X Cell; biotinylated in house N/A Rabbit anti-phospho-Akt (Ser473) (clone D9E) Cell Signaling Technology Cat# 4060; RRID:AB_2315049 Rabbit anti-phospho-p44/42 MAPK (Erk1/ 2, Thr202/Tyr204) (clone 197G2) Cell Signaling Technology Cat# 4377; RRID:AB_331775 Goat anti-rabbit IgG AF647 (polyclonal) Thermo Fisher Scientific Cat# A21245; RRID:AB_2535813 FluoTag-X2 anti-TagFP, AZDye568 NanoTag Biotechnologies Cat# N0502-AF568; RRID:AB_3075935 Bacterial and virus strains Endura electrocompetent cells Biosearch Technologies Cat# 60242-2 Chemicals, peptides, and recombinant proteins AvrII restriction enzyme NEB Cat# R0174S BmtI-HF restriction enzyme NEB Cat# R3658S PmeI restriction enzyme NEB Cat# R0560S PshAI restriction enzyme NEB Cat# R0593S XhoI restriction enzyme NEB Cat# R0146S T4 DNA polymerase NEB Cat# M0203S Q5 Hot Start High-Fidelity DNA polymerase NEB Cat# M0493L (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e1 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific Cat# L3000015 ViralBoost Reagent Alstem Cat# VB100 Lenti-X Concentrator Takara Bio Cat# 631232 Fixable Viability Dye eFluor780 Thermo Fisher Scientific Cat# 65-0865-14 Streptavidin AF647 Thermo Fisher Scientific Cat# S21374 Streptavidin AMCA Vector Laboratories Cat# SA-5008-1 EasySep Streptavidin RapidSpheres StemCell Technologies Cat# 50001 Human CXCL12 (SDF1α) PeproTech Cat# 300-28A S-geranylgeranyl-L-glutathione (GGG) Cayman Chemical; except for Figure 5E for which in house synthesis (Lu et al.28) Cat# 9004116 Bovine serum albumin (BSA), fraction V, fatty acid-free Sigma-Aldrich Cat# 126575 AMPure XP Beads Beckman Coulter Cat# A66380 TransIT-2020 Transfection Reagent VWR Cat# 10767-016 Polybrene Sigma-Aldrich Cat# TR-1003 Coelenterazine 400a NanoLight Cat# 340 Coelenterazine h Nanolight Cat# 301 5-fluorouracil Sigma-Aldrich Cat# F6627 Collagenase VIII from Clostridium histolyticum Sigma-Aldrich Cat# C2139 Mouse serum Sigma-Aldrich Cat# M5905 Doxycycline hyclate Sigma Aldrich Cat# D9891 Tris(2-carboxyethyl)phosphine Fisher Scientific Cat# AC363830010 Pierce protease inhibitor tablet Thermo Fisher Scientific Cat# A32963 Lauryl maltose neopentyl glycol (L-MNG) Anatrace Cat# NG310 ATP Fisher Scientific Cat# AC102800100 Cholesteryl hemisuccinate Steraloids Cat# C6823 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 n-dodecyl-β-d-maltopyranoside (DM) Anatrace Cat# D310 Lambda phosphatase NEB Cat# P0753S Calf intestinal phosphatase NEB Cat# M0290 Antarctic phosphatase NEB Cat# M0289S Leukotriene C4 Cayman Chemical Cat# 20210 S-octane-L-glutathione In house synthesis N/A S-hexadecane-L-glutathione In house synthesis N/A S-isoprenyl-L-glutathione In house synthesis N/A S-farnesyl-L-glutathione In house synthesis N/A Farnesol Sigma-Aldrich Cat# F203 Triphenylphosphine Sigma-Aldrich Cat# T84409 Tetrabromomethane (carbon tetrabromide) Sigma-Aldrich Cat# C11081 N-hexane Sigma-Aldrich Cat# 296090 L-glutathione Sigma-Aldrich Cat# G4251 1-bromooctane Sigma-Aldrich Cat# 152951 3,3-demethylallyl bromide Sigma-Aldrich Cat# 249904 Critical commercial assays Quick Ligation Kit NEB Cat# M2200S Nucleospin Gel & PCR Clean-up Mini Kit Macherey-Nagel Cat# 740609.50 Gibson Assembly Master Mix NEB Cat# E2611S (Continued on next page) e2 Cell Genomics 5, 100981, November 12, 2025 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER QIAquick Gel Extraction Kit Qiagen Cat# 28704 HiFi DNA Assembly Master Mix NEB Cat# E2621S QIAprep Spin Miniprep Kit Qiagen Cat# 27104 HiSpeed Plasmid Midi Prep Kit Qiagen Cat# 12643 Quick-DNA Miniprep Plus Kit Zymo Research Cat# D3025 QIAEX II Gel Extraction Kit Qiagen Cat# 20021 Qubit dsDNA HS Quantification Assay Kit Thermo Fisher Scientific Cat# Q32851 Q5 Site-directed Mutagenesis Kit NEB Cat# E0554S Nextera XT DNA Library Preparation Kit Illumina Cat# FC-131-1096 TapeStation High Sensitivity D5000 Ladder; Reagents; ScreenTape Agilent Cat# 5067–5594; 5067–5593; 5067-5592 RhoA G-LISA GTPase Activation Assay Kit (colorimetric) Cytoskeleton Cat# BK124 Total RhoA ELISA Kit Cytoskeleton Cat# BK150 ExpiFectamine 293 Transfection Kit Thermo Fisher Scientific Cat# A14525 Qubit Protein Assay Kit Thermo Fisher Scientific Cat# Q33211 Deposited data ESM1b human missense variant scores Brandes et al.6 https://huggingface.co/spaces/ntranoslab/ esm_variants gnomAD v4.0.0 Karczewski et al.2 https://gnomad.broadinstitute.org/; RRID: SCR_014964 COSMIC Tate et al.45 https://cancer.sanger.ac.uk/cosmic/login; RRID:SCR_002260 Raw and analyzed P2RY8 DMS data This paper NCBI SRA: PRJNA1179413 (RRID: SCR_004891) GGG-bound P2RY8-miniG13 structure coordinates This paper PDB: 9ECJ; EMD-47912; EMD-47914 Gα13 structure coordinates Chen et al.51 PDB: 7T6B Gβ1γ2 structure coordinates Billesbølle et al.52 PDB: 8F76 GPR68 deep mutational scan fitness score Howard et al.23 MAVEdb: 00001207 Experimental models: Cell lines OCI-Ly8 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_8803 HEK293T Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0063 HEK293T Platinum-E (Plat-E) Retroviral Packaging Line Gift from S.Schwab, NYU Cell Biolabs, Cat# RV101 WEHI-231 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0577 Lenti-X 293T cell Line Takara Bio Cat# 632180 Expi293F TetR inducible cells Thermo Fisher Scientific Cat# A39241 Spodoptera frugiperda Sf9 insect cells Expression Systems Cat# 94-001F Trichoplusia ni Hi5 insect cells Expression Systems Cat# 94-002F Experimental models: Organisms/strains Mouse: C57BL/6J The Jackson Laboratory RRID:IMSR_JAX:000664 Mouse: CD45.1 B6: B6.SJL-Ptprca Pepcb/BoyJ The Jackson Laboratory RRID:IMSR_JAX:002014 (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e3 Article ll OPEN ACCESS
Protease Inhibitor:Article Title: Phenotypic pleiotropy of missense variants in human B cell confinement receptor P2RY8.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human TruStain FcX block Biolegend Cat# 422302; RRID:AB_2818986 InVivoPlus anti-mouse CD16/CD32 (mouse Fc block) Bio X Cell Cat# BE0307; RRID:AB_2736987 Armenian hamster anti-mouse CD3ε biotin (clone 145-2C11) Biolegend Cat# 100303, RRID:AB_312668 Syrian hamster anti-mouse anti-CD3ε AF700 (clone 500A2) Biolegend Cat# 152316; RRID:AB_2632713 Rat anti-mouse CD4 BV711 (clone GK1.5) Biolegend Cat# 100447; RRID:AB_2564586 Rat anti-mouse CD8a BV510 (clone 53-6.7) Biolegend Cat# 100752; RRID:AB_2563057 Rat anti-mouse CD11b BV570 (clone M1/70) Biolegend Cat# 101233; RRID:AB_10896949 Rat anti-mouse CD35 (CR1) biotin (clone 8C12) Fisher Scientific Cat# 553816; RRID:AB_2643602 Mouse anti-mouse CD45.1 APC (clone A20) Cytek Biosciences Cat# 20–0453; RRID:AB_2621575 Mouse anti-mouse CD45.1 PE (clone 104) Biolegend Cat# 109808; RRID:AB_313445 Rat anti-mouse/human CD45R/B220 BV785 (clone RA3-6B2) Biolegend Cat# 103246; RRID:AB_2563256 Hamster anti-mouse CD95 PE-Cy7 (clone Jo2) BD Biosciences Cat# 557653; RRID:AB_396768 Rat anti-mouse CD180 (clone RP/14) BD Biosciences Cat# 562191; RRID:AB_11153851 Rabbit anti-GFP AF488 (polyclonal) Thermo Fisher Scientific Cat# A21311; RRID:AB_221477 Rat anti-GFP AF488 (clone FM2-64G) Biolegend Cat # 338007; RRID:AB_2563287 Rat anti-mouse/human GL7 antigen Pacific Blue (clone GL7) Biolegend Cat# 144614; RRID:AB_2563292 Rat anti-mouse IgD AF647 (clone 11-26c.2a) Biolegend Cat# 405708; RRID:AB_893528 Rat anti-mouse IgD PerCP-Cy5.5 (clone 11-26c.2a) Biolegend Cat# 405710; RRID:AB_1575113 Rat anti-OX56 antigen biotin Contract production Bio X Cell; biotinylated in house N/A Rabbit anti-phospho-Akt (Ser473) (clone D9E) Cell Signaling Technology Cat# 4060; RRID:AB_2315049 Rabbit anti-phospho-p44/42 MAPK (Erk1/ 2, Thr202/Tyr204) (clone 197G2) Cell Signaling Technology Cat# 4377; RRID:AB_331775 Goat anti-rabbit IgG AF647 (polyclonal) Thermo Fisher Scientific Cat# A21245; RRID:AB_2535813 FluoTag-X2 anti-TagFP, AZDye568 NanoTag Biotechnologies Cat# N0502-AF568; RRID:AB_3075935 Bacterial and virus strains Endura electrocompetent cells Biosearch Technologies Cat# 60242-2 Chemicals, peptides, and recombinant proteins AvrII restriction enzyme NEB Cat# R0174S BmtI-HF restriction enzyme NEB Cat# R3658S PmeI restriction enzyme NEB Cat# R0560S PshAI restriction enzyme NEB Cat# R0593S XhoI restriction enzyme NEB Cat# R0146S T4 DNA polymerase NEB Cat# M0203S Q5 Hot Start High-Fidelity DNA polymerase NEB Cat# M0493L (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e1 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific Cat# L3000015 ViralBoost Reagent Alstem Cat# VB100 Lenti-X Concentrator Takara Bio Cat# 631232 Fixable Viability Dye eFluor780 Thermo Fisher Scientific Cat# 65-0865-14 Streptavidin AF647 Thermo Fisher Scientific Cat# S21374 Streptavidin AMCA Vector Laboratories Cat# SA-5008-1 EasySep Streptavidin RapidSpheres StemCell Technologies Cat# 50001 Human CXCL12 (SDF1α) PeproTech Cat# 300-28A S-geranylgeranyl-L-glutathione (GGG) Cayman Chemical; except for Figure 5E for which in house synthesis (Lu et al.28) Cat# 9004116 Bovine serum albumin (BSA), fraction V, fatty acid-free Sigma-Aldrich Cat# 126575 AMPure XP Beads Beckman Coulter Cat# A66380 TransIT-2020 Transfection Reagent VWR Cat# 10767-016 Polybrene Sigma-Aldrich Cat# TR-1003 Coelenterazine 400a NanoLight Cat# 340 Coelenterazine h Nanolight Cat# 301 5-fluorouracil Sigma-Aldrich Cat# F6627 Collagenase VIII from Clostridium histolyticum Sigma-Aldrich Cat# C2139 Mouse serum Sigma-Aldrich Cat# M5905 Doxycycline hyclate Sigma Aldrich Cat# D9891 Tris(2-carboxyethyl)phosphine Fisher Scientific Cat# AC363830010 Pierce protease inhibitor tablet Thermo Fisher Scientific Cat# A32963 Lauryl maltose neopentyl glycol (L-MNG) Anatrace Cat# NG310 ATP Fisher Scientific Cat# AC102800100 Cholesteryl hemisuccinate Steraloids Cat# C6823 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 n-dodecyl-β-d-maltopyranoside (DM) Anatrace Cat# D310 Lambda phosphatase NEB Cat# P0753S Calf intestinal phosphatase NEB Cat# M0290 Antarctic phosphatase NEB Cat# M0289S Leukotriene C4 Cayman Chemical Cat# 20210 S-octane-L-glutathione In house synthesis N/A S-hexadecane-L-glutathione In house synthesis N/A S-isoprenyl-L-glutathione In house synthesis N/A S-farnesyl-L-glutathione In house synthesis N/A Farnesol Sigma-Aldrich Cat# F203 Triphenylphosphine Sigma-Aldrich Cat# T84409 Tetrabromomethane (carbon tetrabromide) Sigma-Aldrich Cat# C11081 N-hexane Sigma-Aldrich Cat# 296090 L-glutathione Sigma-Aldrich Cat# G4251 1-bromooctane Sigma-Aldrich Cat# 152951 3,3-demethylallyl bromide Sigma-Aldrich Cat# 249904 Critical commercial assays Quick Ligation Kit NEB Cat# M2200S Nucleospin Gel & PCR Clean-up Mini Kit Macherey-Nagel Cat# 740609.50 Gibson Assembly Master Mix NEB Cat# E2611S (Continued on next page) e2 Cell Genomics 5, 100981, November 12, 2025 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER QIAquick Gel Extraction Kit Qiagen Cat# 28704 HiFi DNA Assembly Master Mix NEB Cat# E2621S QIAprep Spin Miniprep Kit Qiagen Cat# 27104 HiSpeed Plasmid Midi Prep Kit Qiagen Cat# 12643 Quick-DNA Miniprep Plus Kit Zymo Research Cat# D3025 QIAEX II Gel Extraction Kit Qiagen Cat# 20021 Qubit dsDNA HS Quantification Assay Kit Thermo Fisher Scientific Cat# Q32851 Q5 Site-directed Mutagenesis Kit NEB Cat# E0554S Nextera XT DNA Library Preparation Kit Illumina Cat# FC-131-1096 TapeStation High Sensitivity D5000 Ladder; Reagents; ScreenTape Agilent Cat# 5067–5594; 5067–5593; 5067-5592 RhoA G-LISA GTPase Activation Assay Kit (colorimetric) Cytoskeleton Cat# BK124 Total RhoA ELISA Kit Cytoskeleton Cat# BK150 ExpiFectamine 293 Transfection Kit Thermo Fisher Scientific Cat# A14525 Qubit Protein Assay Kit Thermo Fisher Scientific Cat# Q33211 Deposited data ESM1b human missense variant scores Brandes et al.6 https://huggingface.co/spaces/ntranoslab/ esm_variants gnomAD v4.0.0 Karczewski et al.2 https://gnomad.broadinstitute.org/; RRID: SCR_014964 COSMIC Tate et al.45 https://cancer.sanger.ac.uk/cosmic/login; RRID:SCR_002260 Raw and analyzed P2RY8 DMS data This paper NCBI SRA: PRJNA1179413 (RRID: SCR_004891) GGG-bound P2RY8-miniG13 structure coordinates This paper PDB: 9ECJ; EMD-47912; EMD-47914 Gα13 structure coordinates Chen et al.51 PDB: 7T6B Gβ1γ2 structure coordinates Billesbølle et al.52 PDB: 8F76 GPR68 deep mutational scan fitness score Howard et al.23 MAVEdb: 00001207 Experimental models: Cell lines OCI-Ly8 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_8803 HEK293T Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0063 HEK293T Platinum-E (Plat-E) Retroviral Packaging Line Gift from S.Schwab, NYU Cell Biolabs, Cat# RV101 WEHI-231 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0577 Lenti-X 293T cell Line Takara Bio Cat# 632180 Expi293F TetR inducible cells Thermo Fisher Scientific Cat# A39241 Spodoptera frugiperda Sf9 insect cells Expression Systems Cat# 94-001F Trichoplusia ni Hi5 insect cells Expression Systems Cat# 94-002F Experimental models: Organisms/strains Mouse: C57BL/6J The Jackson Laboratory RRID:IMSR_JAX:000664 Mouse: CD45.1 B6: B6.SJL-Ptprca Pepcb/BoyJ The Jackson Laboratory RRID:IMSR_JAX:002014 (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e3 Article ll OPEN ACCESS
Ligation:Article Title: Phenotypic pleiotropy of missense variants in human B cell confinement receptor P2RY8.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human TruStain FcX block Biolegend Cat# 422302; RRID:AB_2818986 InVivoPlus anti-mouse CD16/CD32 (mouse Fc block) Bio X Cell Cat# BE0307; RRID:AB_2736987 Armenian hamster anti-mouse CD3ε biotin (clone 145-2C11) Biolegend Cat# 100303, RRID:AB_312668 Syrian hamster anti-mouse anti-CD3ε AF700 (clone 500A2) Biolegend Cat# 152316; RRID:AB_2632713 Rat anti-mouse CD4 BV711 (clone GK1.5) Biolegend Cat# 100447; RRID:AB_2564586 Rat anti-mouse CD8a BV510 (clone 53-6.7) Biolegend Cat# 100752; RRID:AB_2563057 Rat anti-mouse CD11b BV570 (clone M1/70) Biolegend Cat# 101233; RRID:AB_10896949 Rat anti-mouse CD35 (CR1) biotin (clone 8C12) Fisher Scientific Cat# 553816; RRID:AB_2643602 Mouse anti-mouse CD45.1 APC (clone A20) Cytek Biosciences Cat# 20–0453; RRID:AB_2621575 Mouse anti-mouse CD45.1 PE (clone 104) Biolegend Cat# 109808; RRID:AB_313445 Rat anti-mouse/human CD45R/B220 BV785 (clone RA3-6B2) Biolegend Cat# 103246; RRID:AB_2563256 Hamster anti-mouse CD95 PE-Cy7 (clone Jo2) BD Biosciences Cat# 557653; RRID:AB_396768 Rat anti-mouse CD180 (clone RP/14) BD Biosciences Cat# 562191; RRID:AB_11153851 Rabbit anti-GFP AF488 (polyclonal) Thermo Fisher Scientific Cat# A21311; RRID:AB_221477 Rat anti-GFP AF488 (clone FM2-64G) Biolegend Cat # 338007; RRID:AB_2563287 Rat anti-mouse/human GL7 antigen Pacific Blue (clone GL7) Biolegend Cat# 144614; RRID:AB_2563292 Rat anti-mouse IgD AF647 (clone 11-26c.2a) Biolegend Cat# 405708; RRID:AB_893528 Rat anti-mouse IgD PerCP-Cy5.5 (clone 11-26c.2a) Biolegend Cat# 405710; RRID:AB_1575113 Rat anti-OX56 antigen biotin Contract production Bio X Cell; biotinylated in house N/A Rabbit anti-phospho-Akt (Ser473) (clone D9E) Cell Signaling Technology Cat# 4060; RRID:AB_2315049 Rabbit anti-phospho-p44/42 MAPK (Erk1/ 2, Thr202/Tyr204) (clone 197G2) Cell Signaling Technology Cat# 4377; RRID:AB_331775 Goat anti-rabbit IgG AF647 (polyclonal) Thermo Fisher Scientific Cat# A21245; RRID:AB_2535813 FluoTag-X2 anti-TagFP, AZDye568 NanoTag Biotechnologies Cat# N0502-AF568; RRID:AB_3075935 Bacterial and virus strains Endura electrocompetent cells Biosearch Technologies Cat# 60242-2 Chemicals, peptides, and recombinant proteins AvrII restriction enzyme NEB Cat# R0174S BmtI-HF restriction enzyme NEB Cat# R3658S PmeI restriction enzyme NEB Cat# R0560S PshAI restriction enzyme NEB Cat# R0593S XhoI restriction enzyme NEB Cat# R0146S T4 DNA polymerase NEB Cat# M0203S Q5 Hot Start High-Fidelity DNA polymerase NEB Cat# M0493L (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e1 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific Cat# L3000015 ViralBoost Reagent Alstem Cat# VB100 Lenti-X Concentrator Takara Bio Cat# 631232 Fixable Viability Dye eFluor780 Thermo Fisher Scientific Cat# 65-0865-14 Streptavidin AF647 Thermo Fisher Scientific Cat# S21374 Streptavidin AMCA Vector Laboratories Cat# SA-5008-1 EasySep Streptavidin RapidSpheres StemCell Technologies Cat# 50001 Human CXCL12 (SDF1α) PeproTech Cat# 300-28A S-geranylgeranyl-L-glutathione (GGG) Cayman Chemical; except for Figure 5E for which in house synthesis (Lu et al.28) Cat# 9004116 Bovine serum albumin (BSA), fraction V, fatty acid-free Sigma-Aldrich Cat# 126575 AMPure XP Beads Beckman Coulter Cat# A66380 TransIT-2020 Transfection Reagent VWR Cat# 10767-016 Polybrene Sigma-Aldrich Cat# TR-1003 Coelenterazine 400a NanoLight Cat# 340 Coelenterazine h Nanolight Cat# 301 5-fluorouracil Sigma-Aldrich Cat# F6627 Collagenase VIII from Clostridium histolyticum Sigma-Aldrich Cat# C2139 Mouse serum Sigma-Aldrich Cat# M5905 Doxycycline hyclate Sigma Aldrich Cat# D9891 Tris(2-carboxyethyl)phosphine Fisher Scientific Cat# AC363830010 Pierce protease inhibitor tablet Thermo Fisher Scientific Cat# A32963 Lauryl maltose neopentyl glycol (L-MNG) Anatrace Cat# NG310 ATP Fisher Scientific Cat# AC102800100 Cholesteryl hemisuccinate Steraloids Cat# C6823 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 n-dodecyl-β-d-maltopyranoside (DM) Anatrace Cat# D310 Lambda phosphatase NEB Cat# P0753S Calf intestinal phosphatase NEB Cat# M0290 Antarctic phosphatase NEB Cat# M0289S Leukotriene C4 Cayman Chemical Cat# 20210 S-octane-L-glutathione In house synthesis N/A S-hexadecane-L-glutathione In house synthesis N/A S-isoprenyl-L-glutathione In house synthesis N/A S-farnesyl-L-glutathione In house synthesis N/A Farnesol Sigma-Aldrich Cat# F203 Triphenylphosphine Sigma-Aldrich Cat# T84409 Tetrabromomethane (carbon tetrabromide) Sigma-Aldrich Cat# C11081 N-hexane Sigma-Aldrich Cat# 296090 L-glutathione Sigma-Aldrich Cat# G4251 1-bromooctane Sigma-Aldrich Cat# 152951 3,3-demethylallyl bromide Sigma-Aldrich Cat# 249904 Critical commercial assays Quick Ligation Kit NEB Cat# M2200S Nucleospin Gel & PCR Clean-up Mini Kit Macherey-Nagel Cat# 740609.50 Gibson Assembly Master Mix NEB Cat# E2611S (Continued on next page) e2 Cell Genomics 5, 100981, November 12, 2025 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER QIAquick Gel Extraction Kit Qiagen Cat# 28704 HiFi DNA Assembly Master Mix NEB Cat# E2621S QIAprep Spin Miniprep Kit Qiagen Cat# 27104 HiSpeed Plasmid Midi Prep Kit Qiagen Cat# 12643 Quick-DNA Miniprep Plus Kit Zymo Research Cat# D3025 QIAEX II Gel Extraction Kit Qiagen Cat# 20021 Qubit dsDNA HS Quantification Assay Kit Thermo Fisher Scientific Cat# Q32851 Q5 Site-directed Mutagenesis Kit NEB Cat# E0554S Nextera XT DNA Library Preparation Kit Illumina Cat# FC-131-1096 TapeStation High Sensitivity D5000 Ladder; Reagents; ScreenTape Agilent Cat# 5067–5594; 5067–5593; 5067-5592 RhoA G-LISA GTPase Activation Assay Kit (colorimetric) Cytoskeleton Cat# BK124 Total RhoA ELISA Kit Cytoskeleton Cat# BK150 ExpiFectamine 293 Transfection Kit Thermo Fisher Scientific Cat# A14525 Qubit Protein Assay Kit Thermo Fisher Scientific Cat# Q33211 Deposited data ESM1b human missense variant scores Brandes et al.6 https://huggingface.co/spaces/ntranoslab/ esm_variants gnomAD v4.0.0 Karczewski et al.2 https://gnomad.broadinstitute.org/; RRID: SCR_014964 COSMIC Tate et al.45 https://cancer.sanger.ac.uk/cosmic/login; RRID:SCR_002260 Raw and analyzed P2RY8 DMS data This paper NCBI SRA: PRJNA1179413 (RRID: SCR_004891) GGG-bound P2RY8-miniG13 structure coordinates This paper PDB: 9ECJ; EMD-47912; EMD-47914 Gα13 structure coordinates Chen et al.51 PDB: 7T6B Gβ1γ2 structure coordinates Billesbølle et al.52 PDB: 8F76 GPR68 deep mutational scan fitness score Howard et al.23 MAVEdb: 00001207 Experimental models: Cell lines OCI-Ly8 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_8803 HEK293T Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0063 HEK293T Platinum-E (Plat-E) Retroviral Packaging Line Gift from S.Schwab, NYU Cell Biolabs, Cat# RV101 WEHI-231 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0577 Lenti-X 293T cell Line Takara Bio Cat# 632180 Expi293F TetR inducible cells Thermo Fisher Scientific Cat# A39241 Spodoptera frugiperda Sf9 insect cells Expression Systems Cat# 94-001F Trichoplusia ni Hi5 insect cells Expression Systems Cat# 94-002F Experimental models: Organisms/strains Mouse: C57BL/6J The Jackson Laboratory RRID:IMSR_JAX:000664 Mouse: CD45.1 B6: B6.SJL-Ptprca Pepcb/BoyJ The Jackson Laboratory RRID:IMSR_JAX:002014 (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e3 Article ll OPEN ACCESS
Polymerase Chain Reaction:Article Title: Phenotypic pleiotropy of missense variants in human B cell confinement receptor P2RY8.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human TruStain FcX block Biolegend Cat# 422302; RRID:AB_2818986 InVivoPlus anti-mouse CD16/CD32 (mouse Fc block) Bio X Cell Cat# BE0307; RRID:AB_2736987 Armenian hamster anti-mouse CD3ε biotin (clone 145-2C11) Biolegend Cat# 100303, RRID:AB_312668 Syrian hamster anti-mouse anti-CD3ε AF700 (clone 500A2) Biolegend Cat# 152316; RRID:AB_2632713 Rat anti-mouse CD4 BV711 (clone GK1.5) Biolegend Cat# 100447; RRID:AB_2564586 Rat anti-mouse CD8a BV510 (clone 53-6.7) Biolegend Cat# 100752; RRID:AB_2563057 Rat anti-mouse CD11b BV570 (clone M1/70) Biolegend Cat# 101233; RRID:AB_10896949 Rat anti-mouse CD35 (CR1) biotin (clone 8C12) Fisher Scientific Cat# 553816; RRID:AB_2643602 Mouse anti-mouse CD45.1 APC (clone A20) Cytek Biosciences Cat# 20–0453; RRID:AB_2621575 Mouse anti-mouse CD45.1 PE (clone 104) Biolegend Cat# 109808; RRID:AB_313445 Rat anti-mouse/human CD45R/B220 BV785 (clone RA3-6B2) Biolegend Cat# 103246; RRID:AB_2563256 Hamster anti-mouse CD95 PE-Cy7 (clone Jo2) BD Biosciences Cat# 557653; RRID:AB_396768 Rat anti-mouse CD180 (clone RP/14) BD Biosciences Cat# 562191; RRID:AB_11153851 Rabbit anti-GFP AF488 (polyclonal) Thermo Fisher Scientific Cat# A21311; RRID:AB_221477 Rat anti-GFP AF488 (clone FM2-64G) Biolegend Cat # 338007; RRID:AB_2563287 Rat anti-mouse/human GL7 antigen Pacific Blue (clone GL7) Biolegend Cat# 144614; RRID:AB_2563292 Rat anti-mouse IgD AF647 (clone 11-26c.2a) Biolegend Cat# 405708; RRID:AB_893528 Rat anti-mouse IgD PerCP-Cy5.5 (clone 11-26c.2a) Biolegend Cat# 405710; RRID:AB_1575113 Rat anti-OX56 antigen biotin Contract production Bio X Cell; biotinylated in house N/A Rabbit anti-phospho-Akt (Ser473) (clone D9E) Cell Signaling Technology Cat# 4060; RRID:AB_2315049 Rabbit anti-phospho-p44/42 MAPK (Erk1/ 2, Thr202/Tyr204) (clone 197G2) Cell Signaling Technology Cat# 4377; RRID:AB_331775 Goat anti-rabbit IgG AF647 (polyclonal) Thermo Fisher Scientific Cat# A21245; RRID:AB_2535813 FluoTag-X2 anti-TagFP, AZDye568 NanoTag Biotechnologies Cat# N0502-AF568; RRID:AB_3075935 Bacterial and virus strains Endura electrocompetent cells Biosearch Technologies Cat# 60242-2 Chemicals, peptides, and recombinant proteins AvrII restriction enzyme NEB Cat# R0174S BmtI-HF restriction enzyme NEB Cat# R3658S PmeI restriction enzyme NEB Cat# R0560S PshAI restriction enzyme NEB Cat# R0593S XhoI restriction enzyme NEB Cat# R0146S T4 DNA polymerase NEB Cat# M0203S Q5 Hot Start High-Fidelity DNA polymerase NEB Cat# M0493L (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e1 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific Cat# L3000015 ViralBoost Reagent Alstem Cat# VB100 Lenti-X Concentrator Takara Bio Cat# 631232 Fixable Viability Dye eFluor780 Thermo Fisher Scientific Cat# 65-0865-14 Streptavidin AF647 Thermo Fisher Scientific Cat# S21374 Streptavidin AMCA Vector Laboratories Cat# SA-5008-1 EasySep Streptavidin RapidSpheres StemCell Technologies Cat# 50001 Human CXCL12 (SDF1α) PeproTech Cat# 300-28A S-geranylgeranyl-L-glutathione (GGG) Cayman Chemical; except for Figure 5E for which in house synthesis (Lu et al.28) Cat# 9004116 Bovine serum albumin (BSA), fraction V, fatty acid-free Sigma-Aldrich Cat# 126575 AMPure XP Beads Beckman Coulter Cat# A66380 TransIT-2020 Transfection Reagent VWR Cat# 10767-016 Polybrene Sigma-Aldrich Cat# TR-1003 Coelenterazine 400a NanoLight Cat# 340 Coelenterazine h Nanolight Cat# 301 5-fluorouracil Sigma-Aldrich Cat# F6627 Collagenase VIII from Clostridium histolyticum Sigma-Aldrich Cat# C2139 Mouse serum Sigma-Aldrich Cat# M5905 Doxycycline hyclate Sigma Aldrich Cat# D9891 Tris(2-carboxyethyl)phosphine Fisher Scientific Cat# AC363830010 Pierce protease inhibitor tablet Thermo Fisher Scientific Cat# A32963 Lauryl maltose neopentyl glycol (L-MNG) Anatrace Cat# NG310 ATP Fisher Scientific Cat# AC102800100 Cholesteryl hemisuccinate Steraloids Cat# C6823 Glyco-diosgenin (GDN) Anatrace Cat# GDN101 n-dodecyl-β-d-maltopyranoside (DM) Anatrace Cat# D310 Lambda phosphatase NEB Cat# P0753S Calf intestinal phosphatase NEB Cat# M0290 Antarctic phosphatase NEB Cat# M0289S Leukotriene C4 Cayman Chemical Cat# 20210 S-octane-L-glutathione In house synthesis N/A S-hexadecane-L-glutathione In house synthesis N/A S-isoprenyl-L-glutathione In house synthesis N/A S-farnesyl-L-glutathione In house synthesis N/A Farnesol Sigma-Aldrich Cat# F203 Triphenylphosphine Sigma-Aldrich Cat# T84409 Tetrabromomethane (carbon tetrabromide) Sigma-Aldrich Cat# C11081 N-hexane Sigma-Aldrich Cat# 296090 L-glutathione Sigma-Aldrich Cat# G4251 1-bromooctane Sigma-Aldrich Cat# 152951 3,3-demethylallyl bromide Sigma-Aldrich Cat# 249904 Critical commercial assays Quick Ligation Kit NEB Cat# M2200S Nucleospin Gel & PCR Clean-up Mini Kit Macherey-Nagel Cat# 740609.50 Gibson Assembly Master Mix NEB Cat# E2611S (Continued on next page) e2 Cell Genomics 5, 100981, November 12, 2025 Article ll OPEN ACCESS .. REAGENT or RESOURCE SOURCE IDENTIFIER QIAquick Gel Extraction Kit Qiagen Cat# 28704 HiFi DNA Assembly Master Mix NEB Cat# E2621S QIAprep Spin Miniprep Kit Qiagen Cat# 27104 HiSpeed Plasmid Midi Prep Kit Qiagen Cat# 12643 Quick-DNA Miniprep Plus Kit Zymo Research Cat# D3025 QIAEX II Gel Extraction Kit Qiagen Cat# 20021 Qubit dsDNA HS Quantification Assay Kit Thermo Fisher Scientific Cat# Q32851 Q5 Site-directed Mutagenesis Kit NEB Cat# E0554S Nextera XT DNA Library Preparation Kit Illumina Cat# FC-131-1096 TapeStation High Sensitivity D5000 Ladder; Reagents; ScreenTape Agilent Cat# 5067–5594; 5067–5593; 5067-5592 RhoA G-LISA GTPase Activation Assay Kit (colorimetric) Cytoskeleton Cat# BK124 Total RhoA ELISA Kit Cytoskeleton Cat# BK150 ExpiFectamine 293 Transfection Kit Thermo Fisher Scientific Cat# A14525 Qubit Protein Assay Kit Thermo Fisher Scientific Cat# Q33211 Deposited data ESM1b human missense variant scores Brandes et al.6 https://huggingface.co/spaces/ntranoslab/ esm_variants gnomAD v4.0.0 Karczewski et al.2 https://gnomad.broadinstitute.org/; RRID: SCR_014964 COSMIC Tate et al.45 https://cancer.sanger.ac.uk/cosmic/login; RRID:SCR_002260 Raw and analyzed P2RY8 DMS data This paper NCBI SRA: PRJNA1179413 (RRID: SCR_004891) GGG-bound P2RY8-miniG13 structure coordinates This paper PDB: 9ECJ; EMD-47912; EMD-47914 Gα13 structure coordinates Chen et al.51 PDB: 7T6B Gβ1γ2 structure coordinates Billesbølle et al.52 PDB: 8F76 GPR68 deep mutational scan fitness score Howard et al.23 MAVEdb: 00001207 Experimental models: Cell lines OCI-Ly8 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_8803 HEK293T Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0063 HEK293T Platinum-E (Plat-E) Retroviral Packaging Line Gift from S.Schwab, NYU Cell Biolabs, Cat# RV101 WEHI-231 Cell line was previously obtained from other laboratories and further authentication was not performed RRID:CVCL_0577 Lenti-X 293T cell Line Takara Bio Cat# 632180 Expi293F TetR inducible cells Thermo Fisher Scientific Cat# A39241 Spodoptera frugiperda Sf9 insect cells Expression Systems Cat# 94-001F Trichoplusia ni Hi5 insect cells Expression Systems Cat# 94-002F Experimental models: Organisms/strains Mouse: C57BL/6J The Jackson Laboratory RRID:IMSR_JAX:000664 Mouse: CD45.1 B6: B6.SJL-Ptprca Pepcb/BoyJ The Jackson Laboratory RRID:IMSR_JAX:002014 (Continued on next page) Cell Genomics 5, 100981, November 12, 2025 e3 Article ll OPEN ACCESS
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