Table of reagents
| Reagent . | Vendor . | Catalog no. . |
|---|---|---|
| Mice | ||
| Wild-type C57BL/6J | Jackson Laboratories | Jax 000664 |
| NeuroD1-Cre (gift from Leiter lab, UMass) | Jackson Laboratories | Jax 028364 |
| TdTomato-GcaMP | Jackson Laboratories | Jax 024477 |
| Villin1-Cre | Jackson Laboratories | Jax 021504 |
| Piezo2-Cre (gift from Patapoutian lab, Scripps Research) | Jackson Laboratories | Jax 027719 |
| Piezo2f/f (gift from Patapoutian lab, Scripps Research) | Jackson Laboratories | Jax 027720 |
| Primers | ||
| Villin forward primer: 5′-TCAAAGGCTCTCTCAC-3′ | Origene | MP218222 |
| Villin reverse primer: 5′-AGCAGTCACCATCGAAGAAGC-3′ | Origene | MP218222 |
| Piezo2 forward primer: 5′-GCACTCTACCTCAGGAAGACTG-3′ | Origene | MP214282 |
| Piezo2 reverse primer: 5′-CAAAGCTGTGCCACCAGGTTCT-3′ | Origene | MP214282 |
| Chromogranin A forward primer: 5′-AGAACCAGAGCCCTGATGCCAA-3′ | Origene | MP202538 |
| Chromogranin A reverse primer: 5′-CTCTGTGGTTGCCTCAAAGCCA-3′ | Origene | MP202538 |
| β-actin forward primer: 5′-GGCTGTATTCCCCTCCATCG-3′ | IDT | |
| β-actin reverse primer: 5′-CCAGTTGGTAACAATGCCATGT-3′ | IDT | |
| HPRT forward primer: 5′-TGGATACAGGCCAGACTTTGTT-3′ | Origene | MP206455 |
| HPRT reverse primer: 5′-CAGATTCAACTTGCGCTCATC-3′ | Origene | MP206455 |
| Antibodies | ||
| Rabbit Piezo2 antibody (IF) | Novus Biologicals | NBP1-78624 |
| Goat 5-HT antibody (IF) | Abcam | ab66047 validated in PMID: 32407674 |
| Goat E-cadherin antibody (IF) | Biotechne | AF748 validated in PMID: 37225711 |
| Rabbit IgG isotype control | Novus Biologicals | NBP2-24891 |
| Goat IgG isotype control | Abcam | ab37373 |
| Donkey anti-Goat IgG (H&L) FITC | Jackson ImmunoResearch | 705-096-147 |
| Donkey anti-Rabbit IgG (H&L) Cy5 | Jackson ImmunoResearch | 711-175-152 |
| Rabbit Piezo2 antibody (co-IP protein detection) | Novus Biologicals | NBP2-61130 |
| Rabbit E-cadherin antibody (co-IP protein detection) | Cell Signaling Technology | 3195 |
| Normal rabbit IgG | Cell Signaling Technology | 2729 |
| Rabbit catenin δ-1 antibody (co-IP protein detection) | Cell Signaling Technology | 59854 |
| Rabbit β-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 8480 |
| Rabbit γ-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 2309 |
| Rabbit α-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 3236 |
| Rabbit vinculin antibody (co-IP protein detection) | Cell Signaling Technology | 13901 |
| Rabbit Pan-actin antibody (co-IP protein detection) | Cell Signaling Technology | 8465 |
| Others (chemicals, media, materials, etc.) | ||
| Capturem IP & Co-IP Kit | Takara Bio | 635721 |
| 12–230 kDa separation module | Thermo Fisher Scientific | NC1461323 |
| 66–440 kDa separation module | Thermo Fisher Scientific | NC1247135 |
| ProteinSimple anti-rabbit detection module | Thermo Fisher Scientific | NC1196891 |
| 35-mm glass bottom dishes | MatTek Corporation | P35G-1.5-14-C |
| Matrigel GFR and Phenol red free basement membrane matrix | Corning | 356231 |
| Dulbecco’s modified eagle’s medium | Sigma-Aldrich | D6429 |
| Bovine serum albumin | Sigma-Aldrich | A7906 |
| Collagenase Type XI | Sigma-Aldrich | C9407 |
| Fetal bovine serum | Thermo Fisher Scientific | 35-011-CV |
| Penicillin/streptomycin | Thermo Fisher Scientific | 221674 |
| L-glutamine | Thermo Fisher Scientific | 25030081 |
| Gentle cell dissociation reagent | StemCell Technologies | 7174 |
| IntestiCult organoid growth medium (Mouse) | StemCell Technologies | 06005 |
| Dulbecco’s phosphate-buffered saline (Ca++, Mg++ free) | StemCell Technologies | 37354 |
| Accell mouse non-target siRNA | Horizon (PerkinElmer) | D-001910 |
| Accell mouse Piezo2 siRNA | Horizon (PerkinElmer) | E-163012 |
| Accell mouse E-cadherin siRNA | Horizon (PerkinElmer) | E-041028 |
| Phalloidin Atto 565 | Sigma-Aldrich | 94072 |
| OCT embedding compound | Sakura Finetek | 4583 |
| UltraCruz blocking reagent | Santa Cruz Biotechnology | 516214 |
| Reagent . | Vendor . | Catalog no. . |
|---|---|---|
| Mice | ||
| Wild-type C57BL/6J | Jackson Laboratories | Jax 000664 |
| NeuroD1-Cre (gift from Leiter lab, UMass) | Jackson Laboratories | Jax 028364 |
| TdTomato-GcaMP | Jackson Laboratories | Jax 024477 |
| Villin1-Cre | Jackson Laboratories | Jax 021504 |
| Piezo2-Cre (gift from Patapoutian lab, Scripps Research) | Jackson Laboratories | Jax 027719 |
| Piezo2f/f (gift from Patapoutian lab, Scripps Research) | Jackson Laboratories | Jax 027720 |
| Primers | ||
| Villin forward primer: 5′-TCAAAGGCTCTCTCAC-3′ | Origene | MP218222 |
| Villin reverse primer: 5′-AGCAGTCACCATCGAAGAAGC-3′ | Origene | MP218222 |
| Piezo2 forward primer: 5′-GCACTCTACCTCAGGAAGACTG-3′ | Origene | MP214282 |
| Piezo2 reverse primer: 5′-CAAAGCTGTGCCACCAGGTTCT-3′ | Origene | MP214282 |
| Chromogranin A forward primer: 5′-AGAACCAGAGCCCTGATGCCAA-3′ | Origene | MP202538 |
| Chromogranin A reverse primer: 5′-CTCTGTGGTTGCCTCAAAGCCA-3′ | Origene | MP202538 |
| β-actin forward primer: 5′-GGCTGTATTCCCCTCCATCG-3′ | IDT | |
| β-actin reverse primer: 5′-CCAGTTGGTAACAATGCCATGT-3′ | IDT | |
| HPRT forward primer: 5′-TGGATACAGGCCAGACTTTGTT-3′ | Origene | MP206455 |
| HPRT reverse primer: 5′-CAGATTCAACTTGCGCTCATC-3′ | Origene | MP206455 |
| Antibodies | ||
| Rabbit Piezo2 antibody (IF) | Novus Biologicals | NBP1-78624 |
| Goat 5-HT antibody (IF) | Abcam | ab66047 validated in PMID: 32407674 |
| Goat E-cadherin antibody (IF) | Biotechne | AF748 validated in PMID: 37225711 |
| Rabbit IgG isotype control | Novus Biologicals | NBP2-24891 |
| Goat IgG isotype control | Abcam | ab37373 |
| Donkey anti-Goat IgG (H&L) FITC | Jackson ImmunoResearch | 705-096-147 |
| Donkey anti-Rabbit IgG (H&L) Cy5 | Jackson ImmunoResearch | 711-175-152 |
| Rabbit Piezo2 antibody (co-IP protein detection) | Novus Biologicals | NBP2-61130 |
| Rabbit E-cadherin antibody (co-IP protein detection) | Cell Signaling Technology | 3195 |
| Normal rabbit IgG | Cell Signaling Technology | 2729 |
| Rabbit catenin δ-1 antibody (co-IP protein detection) | Cell Signaling Technology | 59854 |
| Rabbit β-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 8480 |
| Rabbit γ-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 2309 |
| Rabbit α-catenin antibody (co-IP protein detection) | Cell Signaling Technology | 3236 |
| Rabbit vinculin antibody (co-IP protein detection) | Cell Signaling Technology | 13901 |
| Rabbit Pan-actin antibody (co-IP protein detection) | Cell Signaling Technology | 8465 |
| Others (chemicals, media, materials, etc.) | ||
| Capturem IP & Co-IP Kit | Takara Bio | 635721 |
| 12–230 kDa separation module | Thermo Fisher Scientific | NC1461323 |
| 66–440 kDa separation module | Thermo Fisher Scientific | NC1247135 |
| ProteinSimple anti-rabbit detection module | Thermo Fisher Scientific | NC1196891 |
| 35-mm glass bottom dishes | MatTek Corporation | P35G-1.5-14-C |
| Matrigel GFR and Phenol red free basement membrane matrix | Corning | 356231 |
| Dulbecco’s modified eagle’s medium | Sigma-Aldrich | D6429 |
| Bovine serum albumin | Sigma-Aldrich | A7906 |
| Collagenase Type XI | Sigma-Aldrich | C9407 |
| Fetal bovine serum | Thermo Fisher Scientific | 35-011-CV |
| Penicillin/streptomycin | Thermo Fisher Scientific | 221674 |
| L-glutamine | Thermo Fisher Scientific | 25030081 |
| Gentle cell dissociation reagent | StemCell Technologies | 7174 |
| IntestiCult organoid growth medium (Mouse) | StemCell Technologies | 06005 |
| Dulbecco’s phosphate-buffered saline (Ca++, Mg++ free) | StemCell Technologies | 37354 |
| Accell mouse non-target siRNA | Horizon (PerkinElmer) | D-001910 |
| Accell mouse Piezo2 siRNA | Horizon (PerkinElmer) | E-163012 |
| Accell mouse E-cadherin siRNA | Horizon (PerkinElmer) | E-041028 |
| Phalloidin Atto 565 | Sigma-Aldrich | 94072 |
| OCT embedding compound | Sakura Finetek | 4583 |
| UltraCruz blocking reagent | Santa Cruz Biotechnology | 516214 |