| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| anti-rabbit Piezo1 | Biorbyt | orb41297 |
| anti-rabbit IP3R2 | In house against CPDYRDAQNEGKTVRDGELP | NA |
| Anti-mouse IP3R3 | BD Biosciences | 610312; RRID: AB_397704 |
| anti-rabbit sAC | Thermo Fisher | PA5-25756; RRID: AB_2543256 |
| anti-rabbit Stim1 | Cell Signaling | 4916; RRID: AB_2271287 |
| anti-goat VE-cadherin | Santa Cruz | sc-6458; RRID: AB_2077955 |
| anti-rabbit Total AKT | Cell Signaling | 9272S; RRID: AB_329827 |
| anti-rabbit pAKT (p-Ser 473) | Cell Signaling | 9271T; RRID: AB_329825 |
| anti-mouse β actin | Santa Cruz | sc47778; RRID: AB_626632 |
| anti-mouse α tubulin | Sigma-Aldrich | T6074; RRID: AB_477582 |
| anti-mouse GAPDH | Proteintech | 60004-1-Ig; RRID: AB_2107436 |
| Alexa Fluor 488 Phalloidin | Invitrogen | A12379 |
| anti-mouse GFP | Fisher Scientific | NC9777966 |
| Experimental models: Cell lines | ||
| Human Pulmonary Arterial Endothelial Cells | Lonza | CC-2530, Lot NO:0000662151 CC-2530, Lot NO: 20TL209928 |
| Human Aortic Endothelial Cells | Lonza | CC-2535, Lot NO: 20TL122171 |
| Experimental models: Organisms/strain | ||
| Piezo1 flox/flox | Li et al.18 | NA |
| Cdh5-Cre Piezo1 flox/flox | Friedrich et al.20 | NA |
| Recombinant DNA | ||
| G-CEPIA1er | Suzuki et al.27 | Addgene 58215 |
| mCherry-er | Suzuki et al.27 | NA |
| myr-Akt | Fulton et al.42 | NA |
| AA-Akt | Fulton et al.42 | NA |
| pmaxGFP® Vector | Lonza | VPI-1001 |
| Bacterial and virus strains | ||
| Ad-hVECadp-myr-AKT1 | Vector Biolabs | N/A |
| Ad-GFP | Vector Biolabs | #1060 |
| Chemicals, peptides, and recombinant proteins | ||
| EGM2 | Lonza | CC-4176 |
| Fibronectin | Sigma-Aldrich | F1141 |
| Dextran from Leuconostoc mesenteroides | Millipore-Sigma | D1662 |
| Gene Silencer siRNA Transfection Reagent | Genlantis | T500750 |
| Basic Endothelial Cells Nucleofactor Kit | Lonza | VPI-1001 |
| Fura 2 AM | Life Technologies | F1221 |
| Ionomycin | Life Technologies | I657 |
| MnCl2 | Life Technologies | M1787 |
| Calcium Calibration Buffer Kit | Life Technologies | C3008MP |
| EC media w/o Phenol Red | Cell Applications | 211PR-500 |
| Yoda1 | Sigma-Aldrich | SML1558 |
| Bithionol | Selleck Chemicals | S4552 |
| 1-Oleoyl-2-acetyl-sn-glycerol (OAG) | Sigma-Aldrich | O6754 |
| Thapsigargin | Thermo Fisher | T7458 |
| MINIPULS 3 Peristaltic Pumps | Gilson | |
| 5 m Silicone Tubing 0.8 mm | ibidi | 10841 |
| 1. 6 mm serial connector for μ-Slides | ibidi | 10830 |
| Elbow Luer connector male | ibidi | 10802 |
| μ-Slide I 0.2 | ibidi | 80161 |
| μ-Slide I 0.4 | ibidi | 80171 |
| μ-Slide VI 0.4 | ibidi | 80601 |
| Ripa Buffer | Thermo Fisher | R027 |
| Laemmli Sample Buffer | Bio Rad | 1610747 |
| Critical commercial assays | ||
| Green Up cADDis cAMP Assay Kit | Montana Molecular | U0200G |
| BCA Protein Assay Kit | Thermo Fisher | 23225 |
| Oligonucleotides | ||
| ON-Targe Plus SMART pool Piezo1 siRNA: GCAGCAUGACAGACGACAU UGGAGUAUGCCAACGAGAA UGGCUGAUGUUGUCGACUU GCGCAUCAGUCUACGUUUU |
Horizon Discovery | L-020870-03-0005 |
| ON-Targe Plus SMART pool IP3R2 siRNA: CGAAAUGGCCGCUCUAUUA GCAAUGGGUUGGAGACUAU GGAAUGAAAGGGCAAUUAA GAGCAAUAACUACCGGAUU |
Horizon Discovery | L-006208-02-0005 |
| ITPR3 GCAUGGAGCAGAUCGUGU | Thermo Fischer | 4392420 |
| ADCY10 CCAAAGAACUGGACUCGUA | Horizon Discovery | J-006353-09-0005 |
| Silencer Negative Control siRNA #1 | Thermo Fischer | AM4635 |
| Software and algorithms | ||
| ImageJ (FIJI) | National Institutes of Health | https://fiji.sc/ |
| Prism 7 | Graph Pad | https://www.graphpad.com/scientific-software/prism/ |
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