KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
|
| ||
| Occludin | Invitrogen | 71-1500; RRID: AB_2533977 |
| ZO-1 | Invitrogen | 40-2200; RRID: AB_2533456 |
| E-cadherin | ThermoFisher | 14-3249-82; RRID: AB_1210458 |
| β-Catenin | Invitrogen | PA5-77934; RRID: AB_2735522 |
| c-SrcpY418 | MyBiosource | 190090 |
| AlexaFluor 488-mouse IgG | ThermoFisher | A-11001 |
| FITC-mouse anti-occludin | Invitrogen | 33-1511; RRID: AB_87042 |
| Cy3-rabbit IgG | Millipore Sigma | AP106C; RRID: AB_92245 |
| HRP-mouse IgG | Millipore Sigma | AP127P; RRID: AB_92472 |
| HRP-rabbit IgG | Millipore Sigma | 12-348; RRID: AB_390191 |
| Claudin-3 | Invitrogen | PA5-32353; RRID: AB_2549824 |
| Claudin-5 | Invitrogen | 35-2500; RRID: AB_2533200 |
| Trpv6 | Origene | TA590262 |
| CaV1.3 | Origene | TA326516 |
| β-Actin | Millipore Sigma | A2228; RRID: AB_476697 |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Sodium Chloride | Fisher Scientific | S271-500 |
| Potassium Chloride | Fisher Scientific | P217-500 |
| Magnesium Chloride | Fisher Scientific | BP214-500 |
| Glucose | Sigma | G8270-1KG |
| HEPES | Fisher Scientific | BP310-100 |
| Hydrochloric Acid | Fisher Scientific | A144-212 |
| Sodium Hydroxide | Fisher Scientific | SS266-1 |
| Cesium Chloride | Acros | 7647-17-8 |
| Cesium Hydroxide | Acros | 35103-79-8 |
| EGTA | Fisher Scientific | O2783-100 |
| Lipofectamine 2000 | Invitrogen | 52887 |
| Opti-MEM | Gibco | 31985-070 |
| DMEM | Gibco | 11965-092 |
| Fetal Bovine Serum | Gibco | 10082-147 |
| Penicillin-Streptomycin | Sigma | P4333-100 |
| Ethanol | Fisher Scientific | BP2818-500 |
| Acetaldehyde | Millipore Sigma | 75-07-0 |
| SOR-C13 | Peptide 2.0 Inc. | Custom synthesis |
| BAPTA-AM | Millipore Sigma | 126150-97-8 |
| Indo-1 AM | ThermoFisher | I1223 |
| AlexaFluor 488-phalloidin | Invitrogen | A12379 |
| Hoechst 33342 | Invitrogen | H3570 |
| AlexaFluor 647-dextran | Invitrogen | D22914 |
| pRNATin-H1.2/Neo | Genscript Corp | SD1223 |
| FITC-inulin | Millipore Sigma | F3272 |
| 4-Methylpyrazole | Millipore Sigma | 56010-88-9 |
| N-Acetylcysteine | Millipore Sigma | A7250 |
| Diltiazem | Millipore Sigma | D2521 |
| Lipopolysaccharide (P. aeruginosa) | Millipore Sigma | L9143 |
| Lieber-DeCarli diet | Dyets Inc. | 710260 |
| Maltose dextrin | BioServe | 3650 |
| 3,3-Azo-1-butanol | Santa Cruz Biotech | 25055-82-7 |
| Trypsin | Promega | V5111 |
| Peptide extraction cartridge | Waters | WAT094225 |
| Lipofectamine LTX | ThermoFisher | A12621 |
| Lipofectamine 2000 | ThermoFisher | 11668027 |
| Transwells | Millipore Sigma | CLS3460 |
| Collagenase XI | Millipore Sigma | C7657 |
| Dispase II | Millipore Sigma | D4693 |
| Triton X100 | Millipore Sigma | T8787 |
| Leupeptin | Millipore Sigma | 103476-89-7 |
| Pepstatin-A | Millipore Sigma | 26305-03-3 |
| Aprotinin | Millipore Sigma | A1250000 |
| Bestatin | Millipore Sigma | 65391-42-6 |
| Dithiothreitol | Millipore Sigma | 16096-97-2 |
| Nicotinamide | Millipore Sigma | 1090-61-7 |
| Matrigel | Millipore Sigma | CLS354277 |
| Primocin | InvivoGen | amt-pm-1 |
| Gastrin | Millipore Sigma | G9145 |
| SB202190 | Millipore Sigma | S7067 |
| hHGF | R&D Systems | 2207 |
| R-spondin conditioned medium | Laboratory of Radhakrishna Rao | N/A |
| Noggin conditioned medium | Laboratory of Radhakrishna Rao | N/A |
| Wnt conditioned medium | Laboratory of Radhakrishna Rao | N/A |
| Paraformaldehyde | Millipore Sigma | 30525-89-4 |
| Pluronic F127 | ThermoFisher | P3000MP |
| BODIPY FL-N-(2 aminoethyl)maleimide Pierce-N-Ethylmaleimide | ThermoFisher | B10250 |
| ECL chemiluminescence reagent | ThermoFisher | 34095 |
| RBC lysis buffer | BioLegend | 420301 |
| m-CSF | ThermoFisher | PHC9504 |
| Accutase | Millipore Sigma | A6964 |
| LPS (E. coli) | Millipore Sigma | L4391 |
| A23187 | Millipore Sigma | C7522 |
| Tetramethyl benzidine | Millipore Sigma | T0440 |
| Hydrogen peroxide | Millipore Sigma | 7722-84-1 |
| fMLP | Millipore Sigma | 59880-97-6 |
|
| ||
| Critical commercial assays | ||
|
| ||
| QuikChange Lightning Site-Directed Mutagenesis Kit | Agilent | 210519-5 |
| RT-qPCR kit | Bio Rad | 12012901 |
| Duoset ELISA kit | R&D Systems | DY008 |
| LPS assay kit | ThermoFisher | 88282 |
| AST assay kit | Cayman chemical | 701640 |
| ALT assay kit | Cayman chemical | 700260 |
| Triglyceride assay kit | Pointe Scientific | T7532120 |
| EasySep cell isolation kit | Stem cell technologies | 19762 |
|
| ||
| Experimental models: Cell lines and transgenic mice | ||
|
| ||
| Human: Caco-2bbe | ATCC | CRL-2102 |
| Human: HEK293 | ATCC | CRL-1573 |
| Trpv6−/− mice | N/A | |
| GFP-Ocln/RFP-ZO1 mice | N/A | |
|
| ||
| Oligonucleotides | ||
|
| ||
| See Table S1 for oligonucleotide information | ||
|
| ||
| Recombinant DNA | ||
|
| ||
| Plasmid: TRPV6 pMO | Laboratory of Julio Cordero-Morales | N/A |
| Plasmid: TRPV1 pMO | Laboratory of Julio Cordero-Morales | N/A |
| Plasmid: GFP pMO | Laboratory of Julio Cordero-Morales | N/A |
| Trpv6-shRNA | Laboratory of Radhakrishna Rao | N/A |
|
| ||
| Software and algorithms | ||
|
| ||
| Clampfit 10.4 | Molecular Devices | https://www.moleculardevices.com/ |
| Clampex 10.5 | Molecular Devices | https://www.moleculardevices.com/ |
| OriginPro 2021 | OriginLab | https://www.originlab.com/ |
|
| ||
| Other | ||
|
| ||
| Borosilicate glass | Sutter Instrument | BF150-110-10 |
| Axopatch 200A | Molecular Devices | https://www.moleculardevices.com/ |
| Axon Digidata 1550 | Molecular Devices | https://www.moleculardevices.com/ |
| Individual data deposit | Figshare | [figshare]: [] https://doi.org/10.6084/m9.figshare.13299158x |