Reagents
| Reagent or Resource . | Source . | Clone . | Identifier (RRID/catalog number) . | . |
|---|---|---|---|---|
| Antibodies | ||||
| CD45 | BD Biosciences | 30-F11 | AB_2651134 | |
| CD11b | eBioscience | M1/70 | AB_2896082 | |
| Ly6C | Biolegend | HK1.4 | AB_2562352 | |
| MHCII | Biolegend | M5/114.15.2 | AB_2561397 | |
| F4/80 | Biolegend | BM8 | AB_2562305 | |
| CD36 | BD Biosciences | CRF D-2712 | AB_2739064 | |
| Ly6G | Biolegend | 1A8 | AB_10640452 | |
| Lyve1 | eBioscience | ALY7 | AB_10597449 | |
| CD163 | Biolegend | S15049I | AB_2890710 | |
| CD11c | Biolegend | N418 | AB_830649 | |
| Ednrb | Novus Biologicals | 671917 | AB_3096090 | |
| Folrb | Biolegend | 10/FR2 | AB_2721343 | |
| Hmox1 IF | Abcam | EP1391Y | ab52947 | |
| CD68 IF | Abcam | FA-11 | ab53444 | |
| EDNRB IF | Novus Biologicals | Polyclonal | NLS54 | |
| CD31 IF | R&D Systems | Polyclonal | AF3628 | |
| CD31 | Biolegend | 390 | AB_312900 | |
| CD3 | Biolegend | 17A2 | AB_1732057 | |
| CD4 | Biolegend | GK1.5 | AB_10900241 | |
| PD1 | Biolegend | RMP1-30 | AB_313421 | |
| FOXP3 | eBiosciences | FJK-16 s | AB_914349 | |
| Granzyme B | Biolegend | GB11 | AB_2566333 | |
| InVivoMAb anti-mouse PD-1 (CD279) | Bio X Cell | RMP1-14 | AB_10949053 | |
| InVivoMab anti-mouse CD3 | Bio X Cell | 17A2 | AB_1107630 | |
| CD16/32 Fc Block | BD Biosciences | 2.4G2 | AB_394656 | |
| Chemicals, peptides and recombinant proteins | Sequence | |||
| CTV | Invitrogen | C34557 | ||
| Zombie NIR | Biolegend | 423105 | ||
| Macitentan | MedChemExpress | HY-14184 | ||
| Endothelin 1, human, HiLyte Fluor 488-labeled | AnaSpec | AS-64817-025 | HiLyte Fluor 488-CSCSSLMDKECVYFCHLDIIW (Disulfide Bridge: 1–15 and 3–11) | |
| CD45 MicroBeads, mouse | Miltenyi Biotec | 30F11.1 | 130-052-301 | |
| Anti-F4/80 MicroBeads UltraPure, mouse | Miltenyi Biotec | 130-110-443 | ||
| Pan T Cell Isolation Kit II, mouse | Miltenyi Biotec | 130-095-130 | ||
| Monocyte Isolation Kit (BM), mouse | Miltenyi Biotec | 130-100-629 | ||
| Recombinant Murine M-CSF | PeproTech | 315-02 | ||
| Recombinant Human M-CSF | PeproTech | 300-25 | ||
| TAT-CRE Recombinase | Sigma-Aldrich | SCR508 | ||
| Hemin | Sigma-Aldrich | 51280 | ||
| Trypsin-EDTA (0.25%), phenol red | Gibco | 25200056 | ||
| Dynabeads Mouse T-Activator CD3/CD28 for T-Cell expansion and Activation | Gibco | 11452D | ||
| LS Columns, MACS Cell separation | Miltenyi Biotec | 130-042-401 | ||
| DNase I | Sigma-Aldrich | D4527-200KU | ||
| Collagenase B | Sigma-Aldrich | 11088831001 | ||
| RBC Lysis Buffer, 10X | Santa Cruz Biotechnology | sc-296258 | ||
| Vector Laboratories H-3300 Citrate-Based Antigen Unmasking Solution, pH 6 | Vector Laboratories | H-3300-250 | ||
| Critical commercial assays | ||||
| Iron Assay Kit (Colorimetric) | Abcam | ab83366 | ||
| GenElute Mammalian Total RNA Miniprep Kit with DNAse I digestion | Sigma-Aldrich | RTN70-1KT | ||
| High-Capacity RNA-to-cDNA Kit | Thermo Fisher Scientific | 4387406 | ||
| Experimental models: Cell lines and animal models | ||||
| FS B6PRG | ||||
| UPS | ||||
| Synovial | ||||
| qPCR probes (ThermoFisher Taqman gene expression assays) | Assay ID | |||
| Mouse | ||||
| Ednrb | Mm00432989_m1 | |||
| Arg1 | Mm00475988 m1 | |||
| Ciita | Mm00482914_m1 | |||
| Hmox1 | Mm00516005_m1 | |||
| Slc40a1 | Mm01254822_m1 | |||
| Bach1 | Mm01344527_m1 | |||
| Spic | Mm00488428_m1 | |||
| Hprt | Mm03024075_m1 | |||
| Human | ||||
| EDNRB | Hs00240747_m1 | |||
| SPIC | Hs00745162_s1 | |||
| HPRT | Hs99999909_m1 | |||
| SLC40A1 | Hs00205888_m1 | |||
| Reagent or Resource . | Source . | Clone . | Identifier (RRID/catalog number) . | . |
|---|---|---|---|---|
| Antibodies | ||||
| CD45 | BD Biosciences | 30-F11 | AB_2651134 | |
| CD11b | eBioscience | M1/70 | AB_2896082 | |
| Ly6C | Biolegend | HK1.4 | AB_2562352 | |
| MHCII | Biolegend | M5/114.15.2 | AB_2561397 | |
| F4/80 | Biolegend | BM8 | AB_2562305 | |
| CD36 | BD Biosciences | CRF D-2712 | AB_2739064 | |
| Ly6G | Biolegend | 1A8 | AB_10640452 | |
| Lyve1 | eBioscience | ALY7 | AB_10597449 | |
| CD163 | Biolegend | S15049I | AB_2890710 | |
| CD11c | Biolegend | N418 | AB_830649 | |
| Ednrb | Novus Biologicals | 671917 | AB_3096090 | |
| Folrb | Biolegend | 10/FR2 | AB_2721343 | |
| Hmox1 IF | Abcam | EP1391Y | ab52947 | |
| CD68 IF | Abcam | FA-11 | ab53444 | |
| EDNRB IF | Novus Biologicals | Polyclonal | NLS54 | |
| CD31 IF | R&D Systems | Polyclonal | AF3628 | |
| CD31 | Biolegend | 390 | AB_312900 | |
| CD3 | Biolegend | 17A2 | AB_1732057 | |
| CD4 | Biolegend | GK1.5 | AB_10900241 | |
| PD1 | Biolegend | RMP1-30 | AB_313421 | |
| FOXP3 | eBiosciences | FJK-16 s | AB_914349 | |
| Granzyme B | Biolegend | GB11 | AB_2566333 | |
| InVivoMAb anti-mouse PD-1 (CD279) | Bio X Cell | RMP1-14 | AB_10949053 | |
| InVivoMab anti-mouse CD3 | Bio X Cell | 17A2 | AB_1107630 | |
| CD16/32 Fc Block | BD Biosciences | 2.4G2 | AB_394656 | |
| Chemicals, peptides and recombinant proteins | Sequence | |||
| CTV | Invitrogen | C34557 | ||
| Zombie NIR | Biolegend | 423105 | ||
| Macitentan | MedChemExpress | HY-14184 | ||
| Endothelin 1, human, HiLyte Fluor 488-labeled | AnaSpec | AS-64817-025 | HiLyte Fluor 488-CSCSSLMDKECVYFCHLDIIW (Disulfide Bridge: 1–15 and 3–11) | |
| CD45 MicroBeads, mouse | Miltenyi Biotec | 30F11.1 | 130-052-301 | |
| Anti-F4/80 MicroBeads UltraPure, mouse | Miltenyi Biotec | 130-110-443 | ||
| Pan T Cell Isolation Kit II, mouse | Miltenyi Biotec | 130-095-130 | ||
| Monocyte Isolation Kit (BM), mouse | Miltenyi Biotec | 130-100-629 | ||
| Recombinant Murine M-CSF | PeproTech | 315-02 | ||
| Recombinant Human M-CSF | PeproTech | 300-25 | ||
| TAT-CRE Recombinase | Sigma-Aldrich | SCR508 | ||
| Hemin | Sigma-Aldrich | 51280 | ||
| Trypsin-EDTA (0.25%), phenol red | Gibco | 25200056 | ||
| Dynabeads Mouse T-Activator CD3/CD28 for T-Cell expansion and Activation | Gibco | 11452D | ||
| LS Columns, MACS Cell separation | Miltenyi Biotec | 130-042-401 | ||
| DNase I | Sigma-Aldrich | D4527-200KU | ||
| Collagenase B | Sigma-Aldrich | 11088831001 | ||
| RBC Lysis Buffer, 10X | Santa Cruz Biotechnology | sc-296258 | ||
| Vector Laboratories H-3300 Citrate-Based Antigen Unmasking Solution, pH 6 | Vector Laboratories | H-3300-250 | ||
| Critical commercial assays | ||||
| Iron Assay Kit (Colorimetric) | Abcam | ab83366 | ||
| GenElute Mammalian Total RNA Miniprep Kit with DNAse I digestion | Sigma-Aldrich | RTN70-1KT | ||
| High-Capacity RNA-to-cDNA Kit | Thermo Fisher Scientific | 4387406 | ||
| Experimental models: Cell lines and animal models | ||||
| FS B6PRG | ||||
| UPS | ||||
| Synovial | ||||
| qPCR probes (ThermoFisher Taqman gene expression assays) | Assay ID | |||
| Mouse | ||||
| Ednrb | Mm00432989_m1 | |||
| Arg1 | Mm00475988 m1 | |||
| Ciita | Mm00482914_m1 | |||
| Hmox1 | Mm00516005_m1 | |||
| Slc40a1 | Mm01254822_m1 | |||
| Bach1 | Mm01344527_m1 | |||
| Spic | Mm00488428_m1 | |||
| Hprt | Mm03024075_m1 | |||
| Human | ||||
| EDNRB | Hs00240747_m1 | |||
| SPIC | Hs00745162_s1 | |||
| HPRT | Hs99999909_m1 | |||
| SLC40A1 | Hs00205888_m1 | |||