mouse anti-inactive b1 integrin Search Results


cd29 (stem cell marker)(12g10)  (Biotium)
93
Biotium cd29 (stem cell marker)(12g10)
Cd29 (Stem Cell Marker)(12g10), supplied by Biotium, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/CD29+(Stem+Cell+Marker)(12G10)/custom%40bnc042905-100%40pmc11861568__mmc5
Average 93 stars, based on 1 article reviews
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nano-glo  (Promega)
90
Promega nano-glo
Nano Glo, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/nano+glo/pm39964812-697-197-202
Average 90 stars, based on 1 article reviews
nano-glo - by Bioz Stars, 2026-10
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8d4  (Atlas Antibodies)
93
Atlas Antibodies 8d4
8d4, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/Anti-PJA2/pm39964812-697-115-123
Average 93 stars, based on 1 article reviews
8d4 - by Bioz Stars, 2026-10
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rabbit anti-nanoluc  (Promega)
90
Promega rabbit anti-nanoluc
Rabbit Anti Nanoluc, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/rabbit+anti+nanoluc/pm39964812-697-70-73
Average 90 stars, based on 1 article reviews
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rabbit anti kindlin 2 atlas antibodies cat  (Atlas Antibodies)
93
Atlas Antibodies rabbit anti kindlin 2 atlas antibodies cat
Rabbit Anti Kindlin 2 Atlas Antibodies Cat, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/Anti-FERMT2/pm39964812-697-121-123
Average 93 stars, based on 1 article reviews
rabbit anti kindlin 2 atlas antibodies cat - by Bioz Stars, 2026-10
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rabbit polyclonal anti-egfp  (GeneTex)
90
GeneTex rabbit polyclonal anti-egfp
Rabbit Polyclonal Anti Egfp, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/goat+anti+gfp/pm37330909-154-152-150
Average 90 stars, based on 1 article reviews
rabbit polyclonal anti-egfp - by Bioz Stars, 2026-10
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4705s mouse anti a2 integrin ip wb santa cruz cat  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology 4705s mouse anti a2 integrin ip wb santa cruz cat
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
4705s Mouse Anti A2 Integrin Ip Wb Santa Cruz Cat, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/Integrin+%CE%B12+Antibody/pm39964812-697-91-96
Average 94 stars, based on 1 article reviews
4705s mouse anti a2 integrin ip wb santa cruz cat - by Bioz Stars, 2026-10
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m-mlv reverse transcriptase  (Promega)
90
Promega m-mlv reverse transcriptase
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
M Mlv Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/m+mlv+reverse+transcriptase/pm37330909-154-269-272
Average 90 stars, based on 1 article reviews
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mouse monoclonal anti-paxillin (349  (Becton Dickinson)
90
Becton Dickinson mouse monoclonal anti-paxillin (349
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
Mouse Monoclonal Anti Paxillin (349, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/anti+paxillin/pm37330909-154-106-110
Average 90 stars, based on 1 article reviews
mouse monoclonal anti-paxillin (349 - by Bioz Stars, 2026-10
90/100 stars
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easysep mouse cd4+ t cell isolation kit  (STEMCELL Technologies Inc)
90
STEMCELL Technologies Inc easysep mouse cd4+ t cell isolation kit
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
Easysep Mouse Cd4+ T Cell Isolation Kit, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/easysep+human+t+cell+isolation+kit/pm37330909-154-290-298
Average 90 stars, based on 1 article reviews
easysep mouse cd4+ t cell isolation kit - by Bioz Stars, 2026-10
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rat monoclonal anti-mouse ertr9 igm  (BMA Biomedicals)
90
BMA Biomedicals rat monoclonal anti-mouse ertr9 igm
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
Rat Monoclonal Anti Mouse Ertr9 Igm, supplied by BMA Biomedicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/biotinylated+er+tr9+antibody/pm20041149-203-49-54
Average 90 stars, based on 1 article reviews
rat monoclonal anti-mouse ertr9 igm - by Bioz Stars, 2026-10
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monoclonal antibody mab mca404g  (Bio-Rad)
85
Bio-Rad monoclonal antibody mab mca404g
Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 <t>integrin</t> activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.
Monoclonal Antibody Mab Mca404g, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti-inactive+b1+integrin/Mouse+anti+Human+High+Mobility+Group+Protein+B1/10__1128_slash_cvi__05071___11-94-19-23
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monoclonal antibody mab mca404g - by Bioz Stars, 2026-10
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Image Search Results


Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 integrin activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 1. Design and validation of b1IAS purified extracellular protein (A) Diagram of b1 integrin activation sensor (b1IAS) construct. Created with BioRender.com. (B) Molecular modeling of NanoBiT activity reconstruction in a5b1IAS during hybrid domain swing-out and headpiece opening. The headpiece structures of b1 integrin in closed and open con- formations were modeled from PDB: 7CEB and 7CEB. The structure of NanoBiT was modeled from PDB: 7SNX. (C) The ectodomains of a5b1 WT and a5b1IAS were expressed with a C-terminal disulfide-linked ACID-BASE coiled coil in Expi293F or HEK293 cells. The presence of purified soluble proteins was detected by western blot using anti-a5 (A11) and anti-b1 (TS2/16) in non-reducing conditions. (D) Luminescence of purified ectodomain of a5b1IAS with (clasped) or without (unclasped) the ACID-BASE coiled coil and in the absence (CTL) or presence of immobilized BSA or fibronectin (FN) under resting (Ca2+) or activating (Mn2+) metal ion conditions. Data are mean ± SD of three inde- pendent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: Biomarker Discovery, Activation Assay, Construct, Activity Assay, Western Blot

Figure 2. b1IAS functional characterization in b1/ MEFs (A) Top: western blot showing the expression of b1 integrin and NanoBiT in b1 integrin-null (b1/) mouse embryonic fibroblasts (MEFs) transduced with retroviral vector pLZRS-human WT b1 in- tegrin (b1WT) or -b1IAS. Bottom: fluorescence- activated cell sorting (FACS) analysis using 9EG7 antibody in WT MEFs, b1/, and b1/ trans- duced with b1IAS (b1//b1IAS). (B) Relative adhesion measured by the xCELLi- gence system in WT, b1/, and b1//b1IAS MEFs plated on FN. Data are the mean ± SD of three independent experiments. Statistical anal- ysis: two-way ANOVA and Bonferroni’s post hoc analysis. (C) Confocal microscopy showing vinculin (green) and 9EG7+ active b1 integrins (red) in WT, b1/, and b1//b1IAS MEFs plated on FN. The left im- age insets highlight focal adhesion sites. Scale bar: 20 mm. (D) Luminescent microscopy in the presence of furimazine of b1/ MEFs transfected with either intact NanoLuc (left) or b1IAS (right) and plated on FN. Image insets show how most photons generated by b1IAS localize into bona fide focal adhesion sites (right) compared to the random localization of intact NanoLuc (left). Low-magnifi- cation (left) scale bars: 20 mm; high-magnification (right) scale bars: 5 mm. (E) Luminescence of b1//b1IAS MEFs adhering for 30 min to increasing amounts (125–1,000 ng/ mL) of FN compared to b1/ MEFs transfected with intact NanoLuc (b1//NanoLuc). Data are mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis. (F) Luminescence of b1//b1IAS MEFs adhering for 30 min to FN-coated, increasingly stiff (0.5, 10, and 100 kPa) PAGEs compared to b1/ MEFs transfected with intact NanoLuc (b1//NanoLuc). Data are mean ± SD of three independent exper- iments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 2. b1IAS functional characterization in b1/ MEFs (A) Top: western blot showing the expression of b1 integrin and NanoBiT in b1 integrin-null (b1/) mouse embryonic fibroblasts (MEFs) transduced with retroviral vector pLZRS-human WT b1 in- tegrin (b1WT) or -b1IAS. Bottom: fluorescence- activated cell sorting (FACS) analysis using 9EG7 antibody in WT MEFs, b1/, and b1/ trans- duced with b1IAS (b1//b1IAS). (B) Relative adhesion measured by the xCELLi- gence system in WT, b1/, and b1//b1IAS MEFs plated on FN. Data are the mean ± SD of three independent experiments. Statistical anal- ysis: two-way ANOVA and Bonferroni’s post hoc analysis. (C) Confocal microscopy showing vinculin (green) and 9EG7+ active b1 integrins (red) in WT, b1/, and b1//b1IAS MEFs plated on FN. The left im- age insets highlight focal adhesion sites. Scale bar: 20 mm. (D) Luminescent microscopy in the presence of furimazine of b1/ MEFs transfected with either intact NanoLuc (left) or b1IAS (right) and plated on FN. Image insets show how most photons generated by b1IAS localize into bona fide focal adhesion sites (right) compared to the random localization of intact NanoLuc (left). Low-magnifi- cation (left) scale bars: 20 mm; high-magnification (right) scale bars: 5 mm. (E) Luminescence of b1//b1IAS MEFs adhering for 30 min to increasing amounts (125–1,000 ng/ mL) of FN compared to b1/ MEFs transfected with intact NanoLuc (b1//NanoLuc). Data are mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis. (F) Luminescence of b1//b1IAS MEFs adhering for 30 min to FN-coated, increasingly stiff (0.5, 10, and 100 kPa) PAGEs compared to b1/ MEFs transfected with intact NanoLuc (b1//NanoLuc). Data are mean ± SD of three independent exper- iments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: Functional Assay, Western Blot, Expressing, Transduction, Retroviral, Plasmid Preparation, FACS, Confocal Microscopy, Microscopy, Transfection, Generated

Figure 3. b1IAS functional characterization in genetic b1IAS KI TeloHAECs (A) Western blot showing the expression of b1 in- tegrin and NanoBiT tag in b1IAS knockin (KI) ECs compared to WT TeloHAECs (parental). (B) Relative adhesion measured by the xCELLi- gence system in b1IAS KI ECs plated on FN compared to parental ECs. Data are the mean ± SD of four independent experiments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis. (C) Luminescence of b1IAS KI ECs adhering for 30 min to increasing amounts (125–1,000 ng/mL) of FN, collagen type I (Coll I), or laminin 511 (Lam 511). Data are mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis. (D) Luminescence of b1IAS KI ECs adhering for 30 min to increasing amounts (125–1,000 ng/mL) of vitronectin (VN). Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonferroni’s post hoc anal- ysis. (E) Luminescence of b1IAS KI ECs adhering for 30 min to FN-coated, increasingly stiff (10 and 100 kPa) PAGEs. Data are mean ± SD of four in- dependent experiments. Statistical analysis: two- tailed heteroscedastic Student’s t-test. (F) Luminescence of b1IAS KI ECs adhering for 15 min to 500 ng/mL FN in the presence of 9EG7 and 12G10 antibodies. Data are mean ± SD of three independent experiments. Statistical anal- ysis: one-way ANOVA and Bonferroni’s post hoc analysis. (G) Luminescence of b1IAS KI ECs adhering for 15 min to 500 ng/mL FN in the presence of mAb13 or the pan-av integrin and a5b1 antagonist MK- 0429 (100 mM). Data are mean ± SD of three in- dependent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis. (H) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for TLN1, FERMT2, and FERMT3 and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (I) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for FLRT2, LPHN2, and PLXND1 and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 3. b1IAS functional characterization in genetic b1IAS KI TeloHAECs (A) Western blot showing the expression of b1 in- tegrin and NanoBiT tag in b1IAS knockin (KI) ECs compared to WT TeloHAECs (parental). (B) Relative adhesion measured by the xCELLi- gence system in b1IAS KI ECs plated on FN compared to parental ECs. Data are the mean ± SD of four independent experiments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis. (C) Luminescence of b1IAS KI ECs adhering for 30 min to increasing amounts (125–1,000 ng/mL) of FN, collagen type I (Coll I), or laminin 511 (Lam 511). Data are mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonferroni’s post hoc analysis. (D) Luminescence of b1IAS KI ECs adhering for 30 min to increasing amounts (125–1,000 ng/mL) of vitronectin (VN). Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonferroni’s post hoc anal- ysis. (E) Luminescence of b1IAS KI ECs adhering for 30 min to FN-coated, increasingly stiff (10 and 100 kPa) PAGEs. Data are mean ± SD of four in- dependent experiments. Statistical analysis: two- tailed heteroscedastic Student’s t-test. (F) Luminescence of b1IAS KI ECs adhering for 15 min to 500 ng/mL FN in the presence of 9EG7 and 12G10 antibodies. Data are mean ± SD of three independent experiments. Statistical anal- ysis: one-way ANOVA and Bonferroni’s post hoc analysis. (G) Luminescence of b1IAS KI ECs adhering for 15 min to 500 ng/mL FN in the presence of mAb13 or the pan-av integrin and a5b1 antagonist MK- 0429 (100 mM). Data are mean ± SD of three in- dependent experiments. Statistical analysis: one- way ANOVA and Bonferroni’s post hoc analysis. (H) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for TLN1, FERMT2, and FERMT3 and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (I) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for FLRT2, LPHN2, and PLXND1 and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: Functional Assay, Western Blot, Expressing, Knock-In, Two Tailed Test, Control

Figure 4. siRNA HTS in b1IAS TeloHAECs to identify activators and inhibitors of b1 in- tegrin (A) Schematic of the high-throughput screening (HTS)-selected genes (194), among the 2,970 genes expressed by parental WT TeloHAECs, whose silencing in b1IAS KI ECs induces a decreased luminescent signal (Z score < 2), therefore b1 integrin activators (142, in green), and those whose silencing induces an increased luminescent signal (Z score > 2), therefore b1 in- tegrin inhibitors (52, in red). (B) Bubble plot representing top integrin focused enriched pathways (adjusted p < 0.05) based on candidate genes obtained from the HTS (2 < Z score < 2). All enriched pathways are listed in Table S2. The EnrichR combined score is the log of the p value from the Fisher exact test multiplied by the Z score of the deviation from the expected rank. Bubble color (adjusted p value) was computed using the Benjamini-Hochberg method for correction for multiple hypotheses testing. The gene ratio is the overlap between the input list and the gene sets in each gene set library for ranking a pathway’s relevance to the input list. (C) Luminescent intensity Z score mean of three biological replicates for each endothelial gene whose siRNA was contained in the Qiagen Drug- gable Genome v.3 siRNA library. The listed genes (b1 integrin inhibitors in red and b1 integrin acti- vators in green) were chosen for secondary vali- dation. (D) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for RAP1B, TNS3, and RHOJ and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (E) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for RACGAP1, PJA2, and VEGF-B and then plated on 500 ng/mL FN. Data are mean ± SD of three independent exper- iments. Statistical analysis: one-way ANOVA and Bonferroni’s post hoc analysis.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 4. siRNA HTS in b1IAS TeloHAECs to identify activators and inhibitors of b1 in- tegrin (A) Schematic of the high-throughput screening (HTS)-selected genes (194), among the 2,970 genes expressed by parental WT TeloHAECs, whose silencing in b1IAS KI ECs induces a decreased luminescent signal (Z score < 2), therefore b1 integrin activators (142, in green), and those whose silencing induces an increased luminescent signal (Z score > 2), therefore b1 in- tegrin inhibitors (52, in red). (B) Bubble plot representing top integrin focused enriched pathways (adjusted p < 0.05) based on candidate genes obtained from the HTS (2 < Z score < 2). All enriched pathways are listed in Table S2. The EnrichR combined score is the log of the p value from the Fisher exact test multiplied by the Z score of the deviation from the expected rank. Bubble color (adjusted p value) was computed using the Benjamini-Hochberg method for correction for multiple hypotheses testing. The gene ratio is the overlap between the input list and the gene sets in each gene set library for ranking a pathway’s relevance to the input list. (C) Luminescent intensity Z score mean of three biological replicates for each endothelial gene whose siRNA was contained in the Qiagen Drug- gable Genome v.3 siRNA library. The listed genes (b1 integrin inhibitors in red and b1 integrin acti- vators in green) were chosen for secondary vali- dation. (D) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for RAP1B, TNS3, and RHOJ and then plated on 500 ng/mL FN. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (E) Luminescence of control b1IAS KI ECs (siCTL) and b1IAS KI ECs silenced for RACGAP1, PJA2, and VEGF-B and then plated on 500 ng/mL FN. Data are mean ± SD of three independent exper- iments. Statistical analysis: one-way ANOVA and Bonferroni’s post hoc analysis.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: High Throughput Screening Assay, Control

Figure 5. The E3 ubiquitin ligase PJA2 pro- motes kindlin-2 degradation and inhibits b1 integrin activation in ECs (A) Left: western blot showing the expression of PJA2 in WT TeloHAECs after PJA2 silencing and silenced cells transduced with silencing resistant murine PJA2 (mPJA2). Right: luminescence in- tensity of b1IAS KI ECs plated on 500 ng/mL FN, silenced for PJA2, and rescued with mPJA2. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (B) Western blot showing the expression of kindlin-2, talin-1, and Rap1B in WT TeloHAECs after PJA2 silencing. (C) Left: western blot showing ubiquitinated GFP kindlin-2 pulled down by ubiquitin affinity beads in Phoenix cells overexpressing PJA2 or control construct. The first lane corresponds to the incu- bation of lysate from cells overexpressing control constructs on non-ubiquitinated beads (see STAR Methods). Right: western blot showing the expression of PJA2 in cells used in the ubiquiti- nated assay shown on the left. (D) Confocal microscopy showing 9EG7+ active b1 integrin (green), kindlin-2 (red) and vinculin (blue) in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for PJA2. The image insets highlight focal adhesion sites. Scale bar: 20 mm. (E) Relative maximum Feret diameter (mFD) of adhesion sites (FA) in WT TeloHAECs silenced for PJA2 compared to siCTL. Data are the mean ± SD of three independent experiments (10 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test. (F) Number of adhesion sites (FA) in siCTL and siPJA2 ECs as in (E). Data are the mean ± SD of three independent experiments (9 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 5. The E3 ubiquitin ligase PJA2 pro- motes kindlin-2 degradation and inhibits b1 integrin activation in ECs (A) Left: western blot showing the expression of PJA2 in WT TeloHAECs after PJA2 silencing and silenced cells transduced with silencing resistant murine PJA2 (mPJA2). Right: luminescence in- tensity of b1IAS KI ECs plated on 500 ng/mL FN, silenced for PJA2, and rescued with mPJA2. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (B) Western blot showing the expression of kindlin-2, talin-1, and Rap1B in WT TeloHAECs after PJA2 silencing. (C) Left: western blot showing ubiquitinated GFP kindlin-2 pulled down by ubiquitin affinity beads in Phoenix cells overexpressing PJA2 or control construct. The first lane corresponds to the incu- bation of lysate from cells overexpressing control constructs on non-ubiquitinated beads (see STAR Methods). Right: western blot showing the expression of PJA2 in cells used in the ubiquiti- nated assay shown on the left. (D) Confocal microscopy showing 9EG7+ active b1 integrin (green), kindlin-2 (red) and vinculin (blue) in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for PJA2. The image insets highlight focal adhesion sites. Scale bar: 20 mm. (E) Relative maximum Feret diameter (mFD) of adhesion sites (FA) in WT TeloHAECs silenced for PJA2 compared to siCTL. Data are the mean ± SD of three independent experiments (10 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test. (F) Number of adhesion sites (FA) in siCTL and siPJA2 ECs as in (E). Data are the mean ± SD of three independent experiments (9 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: Ubiquitin Proteomics, Activation Assay, Western Blot, Expressing, Transduction, Control, Construct, Confocal Microscopy, Two Tailed Test

Figure 6. VEGF-B is an effective inhibitor of b1 integrin activation in ECs (A) Left: western blot showing the expression of VEGF-B protein in WT TeloHAECs after VEGFB silencing compared to siCTL. Right: lumines- cence of TeloHAEC b1IAS plated on 500 ng/mL FN and silenced for VEGFB in the presence or absence of exogenous VEGF-B for 15 min. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (B) Confocal microscopy showing 9EG7+ active b1 integrin (green), kindlin-2 (red), and vinculin (blue) in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for VEGFB. The image insets highlight focal adhesion sites. Scale bar: 20 mm. (C) Relative maximum Feret diameter (mFD) of adhesion sites (FA) in WT TeloHAECs silenced for VEGFB compared to siCTL. Data are the mean ± SD of three independent experiments (10 cells each). Statistical analysis: two-tailed hetero- scedastic Student’s t test. (D) Number of adhesion sites (FA) in WT Telo- HAECs silenced for VEGFB compared to siCTL. Data are the mean ± SD of three independent experiments (9 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test. (E) Relative adhesion measured by the xCELLigence system in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for VEGFB. Data are the mean ± SD of five independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis. (F) Relative adhesion measured by the xCELLi- gence system in WT TeloHAECs plated on 1.5 mg/mL FN and treated or not with 100 and 200 ng/mL exogenous VEGF-B. Data are the mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis.

Journal: Cell reports

Article Title: Luminescent sensing of conformational integrin activation in living cells.

doi: 10.1016/j.celrep.2025.115319

Figure Lengend Snippet: Figure 6. VEGF-B is an effective inhibitor of b1 integrin activation in ECs (A) Left: western blot showing the expression of VEGF-B protein in WT TeloHAECs after VEGFB silencing compared to siCTL. Right: lumines- cence of TeloHAEC b1IAS plated on 500 ng/mL FN and silenced for VEGFB in the presence or absence of exogenous VEGF-B for 15 min. Data are mean ± SD of three independent experiments. Statistical analysis: one-way ANOVA and Bonfer- roni’s post hoc analysis. (B) Confocal microscopy showing 9EG7+ active b1 integrin (green), kindlin-2 (red), and vinculin (blue) in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for VEGFB. The image insets highlight focal adhesion sites. Scale bar: 20 mm. (C) Relative maximum Feret diameter (mFD) of adhesion sites (FA) in WT TeloHAECs silenced for VEGFB compared to siCTL. Data are the mean ± SD of three independent experiments (10 cells each). Statistical analysis: two-tailed hetero- scedastic Student’s t test. (D) Number of adhesion sites (FA) in WT Telo- HAECs silenced for VEGFB compared to siCTL. Data are the mean ± SD of three independent experiments (9 cells each). Statistical analysis: two-tailed heteroscedastic Student’s t test. (E) Relative adhesion measured by the xCELLigence system in WT TeloHAECs plated on 1.5 mg/mL FN and silenced for VEGFB. Data are the mean ± SD of five independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis. (F) Relative adhesion measured by the xCELLi- gence system in WT TeloHAECs plated on 1.5 mg/mL FN and treated or not with 100 and 200 ng/mL exogenous VEGF-B. Data are the mean ± SD of three independent experiments. Statistical analysis: two-way ANOVA and Bonfer- roni’s post hoc analysis.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti a-tubulin Sigma Cat#T5168; Clone: B-5-1-2; RRID:AB_477579 Mouse anti-vinculin Sigma Cat#V9131; Clone: hVIN-1; RRID:AB_477629 Mouse anti-TNS3 Sigma Cat#SAB4200416; Clone: TN-17 Mouse anti-b1 integrin Bio Legend Cat#303010; Clone: TS2/16; RRID:AB_314326 Mouse anti-b1 integrin Abcam Cat#ab30388; Clone: JB1B; RRID:AB_775736 Rat anti-active b1 integrin BD Bioscience Cat#553715; Clone: 9EG7; RRID:AB_395001 Mouse anti-b1 integrin Sigma Cat# MAB2247; Clone: 12G10 Mouse anti-inactive b1 integrin EMD Millipore Cat#MABT821; Clone: mAb13 Rabbit anti-NanoLuc Promega Gift from Promega Mouse anti-a5 integrin (IP) BD Pharmingen Cat#555650; RRID:AB_2233951 Rabbit anti-a5 integrin (WB) Cell Signaling Cat4705S Mouse anti-a2 integrin (IP/WB) Santa Cruz Cat#sc-74466; Clone: C-9; RRID:AB_1124939 Rabbit anti- av integrin (IP/WB) Abcam Cat#ab124968; RRID:AB_11129746 Mouse anti-Talin-1 Abcam Cat#ab157808; Clone: 8D4; RRID:AB_3076684 Rabbit anti-PJA2 Abcam Cat#ab272665 Rabbit anti-kindlin-2 Atlas antibodies Cat#HPA040505; RRID:AB_10673300 Mouse anti-kindlin-3 Invitrogen Cat#PA5-26932 Rabbit anti-Rap1B Invitrogen Cat#PA5-29396; RRID:AB_2546872 Mouse anti-RhoJ Invitrogen Cat# MA5-26143; RRID:AB_2725234 Mouse anti-RacGAP1 Santa Cruz Cat#sc-271110; Clone: A-6; RRID:AB_10611939 Mouse anti-VEGF-B Santa Cruz Cat#sc-80442; Clone: J-14I; RRID:AB_1131225 Rabbit anti-GFP antibody Invitrogen Cat#A-11122; RRID:AB_221569 Goat HRP-conjugated anti-mouse Jackson Research Cat#115-035-003; RRID:AB_10015289 Goat HRP-conjugated anti-rabbit Santa Cruz Cat#sc-2054; RRID:AB_631748 Anti-Rat IgG-488 Thermo Fisher Cat#A21470; RRID:AB_10561519 Anti-Rabbit IgG-555 Thermo Fisher Cat#A21428; RRID:AB_2535849 Anti-Mouse IgG-647 Thermo Fisher Cat#A21463; RRID:AB_2535869 Chemicals, peptides, and recombinant proteins CellTiter-Glo Promega G7571 Nano-Glo Promega N1110 Fluorescence Mounting Medium DAKO S3023 Dithiothreitol (DTT) Sigma D0632 Idodacetamide (IAA) Sigma I6125 Endopeptidase Lyc New England Biolabs Neb#P8101 Acrylamide National Diagnostics EC-890 Human Fibronectin R&D Systems 1918-FN Human placental collagen type I Sigma C774 Human placental laminin 511 Gift by Lydia Sorokin (Institute of Physiological Chemistry and Pathobiochemistry, University of M€unster, M€unster, Germany) Previously characterized and validated in Sixt et al.30 Human VEGF-B R&D Systems 751-VE Poly-D-Lysine Gibco A3890401 (Continued on next page) Cell Reports 44, 115319, February 25, 2025 15

Techniques: Activation Assay, Western Blot, Expressing, Confocal Microscopy, Two Tailed Test