KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
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| ||
| guinea pig anti insulin 1:5 | Dako | Cat# IR002; RRID:AB_2800361 |
| Goat anti-Pdx1 1:2500 | Generous gift from Chris Wright | |
| Rabbit anti-Nkx6.1 1:100 | Abcam | Cat# Ab221549; RRID:AB_2754979 |
| Rabbit monoclonal anti Ki-67 | Thermo Fisher Scientific | Cat# RM-9106; RRID:AB_2341197 |
| Mouse anti-glucagon | Abcam | Cat# Ab10988; RRID:AB_297642 |
| Mouse monoclonal anti BrdU (5-bromo-2-deoxyuridine) | GE Healthcare Life Biosciences | Cat# RPN202; RRID:AB_2314032 |
| Goat anti-GFP 1:400 | Abcam | Cat# AB6673; RRID:AB_305643 |
| Rabbit anti Somatostatin 1:200 | Abcam | Cat# Ab108456; RRID:AB_11158517 |
| Alexa Fluor® 488 AffiniPure Donkey Anti-Rabbit IgG (H + L) | Jackson ImmunoResearch Labs | Cat# 711–545; RRID:AB_2313584 |
| Alexa Fluor® 488 AffiniPure Donkey Anti-Goat IgG (H + L) | Jackson ImmunoResearch Labs | Cat# 705–545; RRID:AB_2336933 |
| Alexa Fluor 488-AffiniPure Donkey Anti-Guinea Pig IgG (H + L) | Jackson ImmunoResearch Labs | Cat# 706–545; RRID:AB_2340472 |
| Cy3-AffiniPure Donkey Anti-Mouse IgM, μ Chain Specific | Jackson ImmunoResearch Labs | Cat# 715–165; RRID:AB_2340812 |
| Alexa Fluor 647-AffiniPure Donkey Anti-Mouse IgM, μ Chain Specific | Jackson ImmunoResearch Labs | Cat# 715–605; RRID:AB_2340863 |
| Cy3-AffiniPure Donkey Anti-Rabbit IgG (H + L) | Jackson ImmunoResearch Labs | Cat# 711–165; RRID:AB_2307443 |
| Cy5-AffiniPure Donkey Anti-Rabbit IgG (H + L) | Jackson ImmunoResearch Labs | Cat# 711–175; RRID:AB_2340607 |
|
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| Biological samples | ||
|
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| Pancreatic islets isolated from WT ICR mice for ex vivo experiments | Envigo, Israel | Cat# HSD:ICR MICE - 260 |
|
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| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Ethidium Homodimer III- Dead cell stain 1:500 | Biotum | Cat# 40050 |
| DAPI | Roche | Cat# 10236276001 |
| CAS-Block™ | Thermo Fisher Scientific | Cat# 008120 |
| Tamoxifen (20mg/mL in corn oil) | Sigma | Cat# T5648 |
| Doxycycline hydrochloride, (200μg/mL, 2% w/v sucrose) | Proteogenix | Cat# A600889–0100 |
| Insulin pellets | LinBit implants LinShin, Scarborough ON, Canada | Cat# Pr-1-B |
| Dapagliflozin 25mg/kg/day in drinking water | AstraZeneca | |
| Ketogenic diet | Envigo, Israel | Cat# TD. 96355 |
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| Critical commercial assays | ||
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| ||
| In Situ Cell Death Detection Kit, Fluorescein (TUNEL) | Roche | Cat# 11684795910 |
| QIAprep Spin Miniprep Kit | QIAGEN | Cat# 27106 |
| RNeasy Mini Kit | QIAGEN | Cat# 74106 |
|
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| Deposited data | ||
|
| ||
| RNA-Seq dataset of WT mouse islets exposed to 11.8, 20 and 35mM glucose for 72 h | This paper | GSE209751 |
| RNA-Seq dataset of EndoC-BetaH2 exposed to 5, 20 and 35mM glucose for 72 h | This paper | GSE212749 |
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| Experimental models: Cell lines | ||
|
| ||
| EndoC-betaH2 – human pancreatic beta cells | provided by Endocell and Raphael Scharfmann | https://doi.org/10.1172/JCI72674 |
|
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| Experimental models: Organisms/strains | ||
|
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| Insulin-rtTA; TET-DTA | https://www.zotero.org/google-docs/?NmVJ8n | N/A |
| MipCreER | https://www.zotero.org/google-docs/?ePSfVp | N/A |
| Rosa26-LSL-YFP | https://www.zotero.org/google-docs/?VgU7i3 | N/A |
|
| ||
| Oligonucleotides | ||
|
| ||
| Insulin-CreER_F 5′-TGCCACGACCAAGTGACAGC-3′ | Dor et al.47 | N/A |
| Insulin-CreER_R 5′-CCAGGTTACGGATATAGTTCATG-3′ | Dor et al.47 | N/A |
| ROSA26_F 5′-AAAGTCGCTCTGAGTTGTTAT-3′ | Soriano et al.48 | N/A |
| WT 5′-GAAAGACCGCGAAGAGTTTG-3′ | Soriano et al.48 | N/A |
| ROSA26_R 5′-TAAGCCTGCCCAGAAGACTC-3′ | Soriano et al.48 | N/A |
| TET-DTA_F 5′-TTTTGACCTCCATAGAAGAC-3′ | Lee et al.49 | N/A |
| TET-DTA_R 5′-GGCATTATCCACTTTTAGTGC-3′ | Lee et al.49 | N/A |
| Insulin-rtTA_F 5′-TAGATGTGCTTTACTAAGTCATCGCG-3′ | Milo-Landesman et al.50 | N/A |
| Insulin-rtTA_R 5′-AGATCGAGCAGGCCCTCGATGGTAG-3′ | Milo-Landesman et al.50 | N/A |
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| Software and algorithms | ||
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| NIS-Elements | Nikon | RRID:SCR_014329 |
| GraphPad Prism 7 | GraphPad | https://www.graphpad.com/ |
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| Adobe Photoshop CC | Adobe | http://www.adobe.com/es/products/photoshop.html |