| Antibodies |
|
| anti-CD3 |
BD |
Cat#553057: RRID: AB_394590 |
| anti-CD28 |
BD |
Cat#553294: RRID:AB_394763 |
| anti-F4/80 - FITC |
Biolegend |
Cat# 123107: RRID:AB_893500 |
| anti-CD11c |
BD |
Cat# 561241: RRID:AB_10611727 |
| anti-IL-1 beta (Pro-form) monoclonal antibody |
Invitrogen |
Cat# 12-7114-80: RRID:AB_10736472 |
| anti-TNF-α Antibody |
Biolegend |
Cat# 506314: RRID:AB_493330 |
| anti-MHC-II |
Biolegend |
Cat# 107616: RRID:AB_493523 |
| anti-CD80 |
Biolegend |
Cat# 104718: RRID:AB_492824 |
| anti-CD86 |
Biolegend |
Cat# 105008: RRID:AB_313151 |
| anti-CD4-FITC |
Biolegend |
Cat# 100510: RRID:AB_312713 |
| anti-CD8 |
Biolegend |
Cat# 100734,:RRID:AB_2075238 |
| anti-IL17A-AlexaFluor647 |
BD |
Cat# 560184: RRID:AB_1645204 |
| anti-IFNg-PE |
BD |
Cat# 554412: RRID:AB_395376 |
| anti- FoxP3-APC |
BD |
Cat# 560401: RRID:AB_1645201 |
| anti-Insulin |
Agilent Dako |
Cat# IR002: RRID:AB_2800361 |
| anti-Insulin |
ProteinTech |
Cat#66198: RRID:AB_2883281 |
| anti-CD3 |
Abcam |
Cat# ab16669: RRID:AB_443425 |
| anti-B220 |
Biolegend |
Cat# 103201: RRID:AB_312986 |
| anti-phospo-eIF2a |
Abcam |
Cat# ab32157: RRID:AB_732117 |
| anti-G3BP1 |
Novus |
Cat# NBP2-16563 |
| anti-PDL1 |
Abcam |
Cat# ab213480: RRID:AB_2773715 |
| anti-PDL1 |
Cell Signaling |
Cat# 29122: RRID:AB_2798970 |
| anti-Actin |
Cell Signaling |
Cat# 3700: RRID:AB_2242334 |
| anti-PDL1 |
BioXCell |
Cat# BE0101: RRID:AB_10949073 |
| anti-IgG |
BioXCell |
Cat# BE0090: RRID:AB_1107780 |
| Alexa Fluor 568 donkey anti-rabbit antibody |
Invitrogen |
Cat# A10042: RRID:AB_2534017 |
| Alexa Fluor 488 donkey anti-guinea-pig antibody |
Invitrogen |
Cat# A-11055: RRID:AB_2534102 |
| anti-IL-6 |
Fluidigm |
3167003C |
| anti-IL-4 |
Fluidigm |
3166003C |
| anti-IL-10 |
Fluidigm |
3158002C |
| anti-TNFa |
Fluidigm |
3162002C |
| anti-IFNg |
Fluidigm |
3165003C |
| anti-IL-17A |
Fluidigm |
3174002C |
| anti-CD4 |
Fluidigm |
3145002C |
| anti-CD8a |
Fluidigm |
3168003C |
| anti-CD11c |
Fluidigm |
3142003C |
| anti-CD206 (MMR) |
Fluidigm |
3169021C |
| anti-I-A/I-E |
Fluidigm |
3209006C |
| anti-CD86 |
Fluidigm |
3172016C |
| anti-CD25 (IL-2R) |
Fluidigm |
3151007C |
| anti-Ly-6C |
Fluidigm |
3150010C |
| anti-CD19 |
Fluidigm |
3149002C |
| anti-CD3e |
Fluidigm |
3152004C |
| anti-CD45 |
Fluidigm |
3089005C |
| anti-F4/80 |
Fluidigm |
3146008C |
| anti-PDL1 |
Fluidigm |
3153016C |
| anti-PD1 |
Fluidigm |
3159024C |
| anti-MHC-I |
Fluidigm |
3143016C |
| anti-CTLA4 |
Fluidigm |
3154008C |
| anti-iNOS |
Fluidigm |
3161011C |
| anti-CD11b |
Fluidigm |
3143015C |
| anti-CD80 |
Fluidigm |
3171008C |
| anti-IL-6 |
Fluidigm |
3167003C |
| anti-IL-4 |
Fluidigm |
3166003C |
| anti-IL-10 |
Fluidigm |
3158002C |
| anti-TNFa |
Fluidigm |
3162002C |
|
| Biological samples |
|
| Human Islets |
IIDP; Scharp-Lacy |
RRID:SAMN19470079 |
| Human Islets |
IIDP; So CA Islet Cell Resource Center |
RRID:SAMN19591106 |
| Human Islets |
IIDP; University of Pennsylvania |
RRID:SAMN19897466 |
| Human Islets |
University of Alberta Islet Core |
RRID:SAMN19859645 |
| Human Islets |
University of Alberta Islet Core |
RRID:SAMN19796386 |
|
| Chemicals, peptides, and recombinant proteins |
|
| RPMI Medium 1640 (1X) |
Gibco |
11875-093 |
| PBS |
Gibco |
14190-144 |
| HEPES Buffer |
Corning |
25-060-CI |
| Penicillin Streptomycin |
Thermo Fisher Scientific |
15140-122 |
| Heat inactivated FBS |
Gibco |
10082-147 |
| EDTA |
Thermo Fisher Scientific |
BP120-500 |
| Xylene |
Thermo Fisher Scientific |
X5-500 |
| Methanol |
Thermo Fisher Scientific |
A412-500 |
| Ethanol |
Thermo Fisher Scientific |
A412-500 |
| Unmasking solution |
Vector labs |
H3300 |
| Horse blocking buffer |
Vector labs |
S-2012-50 |
| Immpress reagent kit peroxidase conjugated anti-rabbit Ig |
Vector labs |
MP-7402 |
| Vector DAB peroxidase substrate kit |
Vector labs |
SK-4100 |
| Hematoxilin |
Sigma Aldrich |
GHS216 |
| Permount |
Fisher Scientific |
12-545M |
| Accutase |
EMD Millipore Corporation |
SCR005 |
| Collagenase |
Sigma Aldrich |
C7657 |
| DNAse I |
ThermoFisher |
AM222 |
| eBioscience™ 1X RBC Lysis Buffer |
Invitrogen |
00-4333-57 |
| DAPI |
Thermo Fisher Scientific |
D1306 |
| Formaldehyde |
Fisher Scientific |
04042-500 |
| Tamoxifen |
Sigma |
T5648 |
| ISRIB |
Sigma |
SML0843 |
| Recombinant IFN-γ |
Prospec |
Cyt-358-b |
| Recombinant IL-1β |
Prospec |
Cyt-272-b |
| Recombinant TNF-α |
Prospec |
Cyt-252-b |
| Recombinant Mouse IL-2 (carrier-free) |
Biolegend |
575404 |
| Recombinant Mouse IL-12 (p70) (carrier-free) |
Biolegend |
577004 |
|
| Critical commercial assays |
|
| High-Capacity cDNA Reverse Transcription Kit |
Applied Biosystems |
4368814 |
| EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit |
Stemcell technologies |
19765 |
| eBioscience™ Foxp3 / Transcription Factor Staining Buffer Set |
eBioscience |
00-5523-01 |
| Golgi stop |
BD |
512092KZ |
| Ionomycin |
Sigma Aldrich |
I0634 |
| Phorbol 12-myristate 13-acetate |
Sigma Aldrich |
P 8139 |
| Cell-IDTM Cisplatin |
Fluidigm |
201064 |
| Cell-ID Intercalator-Ir — 125 μM |
Fluidigm |
201192A |
| Maxpar® Cell Staining Buffer |
Fluidigm |
201068 |
| Maxpar Fix and Perm Buffer |
Fluidigm |
201067 |
| Maxpar Fix I Buffer (5X) |
Fluidigm |
201065 |
| Maxpar Perm-S Buffer |
Fluidigm |
201066 |
| Maxpar PBS |
Fluidigm |
201058 |
| Maxpar Cell Acquisition Solution |
Fluidigm |
201237 |
| Maxpar Water |
Fluidigm |
201069 |
| Chromium Single Cell 3′ Reagent Kits V3 |
10× Genomics |
PN-1000075, PN-1000073, PN-120262 |
| Live/Dead Viability/Cytotoxicity |
Invitrogen |
L3224 |
|
| Deposited data |
|
| RNA Sequencing data |
GEO |
Accession: GSE166572
|
|
| Experimental models: Organisms/strains |
|
| B6.Cg-Alox15tm1.1Nadl/J |
Jackson Laboratories |
RRID:IMSR_JAX:031835 |
| Pdx1PB-CreERTM
|
(Zhang et al., 2005) |
N/A |
| B6.129S2-Alox15/J |
Jackson Laboratories |
RRID:IMSR_JAX:002778 |
| NOD/ShiLtJ |
Jackson Laboratories |
RRID:IMSR_JAX:001976 |
|
| Oligonucleotides |
|
| See Table S3
|
N/A |
N/A |
|
| Software and algorithms |
|
| Cytobank |
Beckman Coulter |
https://www.cytobank.org/
|
| FlowJo |
FlowJo |
10.7 |
| Fiji |
ImageJ |
Version 2.0.0-rc-69/1.52p |
| GraphPad Prism |
GraphPad software, Inc. |
Version 9.0.1 |
| CV-x |
Keyence |
http://www.keyence.com
|
| CellRanger 3.0.2 |
(Butler et al., 2018; Stuart et al., 2019) |
http://support.10xgenomics.com/
|
| R package Seurat |
(Butler et al., 2018; Stuart et al., 2019) |
Version 3.0.0.9200 |
| R package scatter |
(McCarthy et al., 2017) |
N/A |
| R package muscat |
(Crowell et al., 2020) |
Version 1.0.1 |
| EdgeR |
(Robinson et al., 2010) |
N/A |
