KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| CD28 antibody | Thermo Fisher Scientific | Cat # 16-0289-85; RRID: AB_468927 |
| PI3 Kinase p85 antibody | Cell Signaling Technology | Cat # 4292; RRID: AB_329869 |
| Alexa Fluor 647 CD3ε antibody | BioLegend | Cat # 317312; RRID: AB_571883 |
| Pacific Blue PD-1 antibody | BioLegend | Cat # 329915; RRID: AB_1877194 |
| PE/Cy7 PD-L1 antibody | BioLegend | Cat # 329717; RRID: AB_2561686 |
| ZAP70 pY493 antibody | Cell Signaling Technology | Cat # 2704S; RRID: AB_2217457 |
| PE mouse PD-1 antibody | BioLegend | Cat # 109103; RRID: AB_313420 |
| APC mouse PD-1 antibody | BioLegend | Cat # 109111; RRID: AB_10613470 |
| PE mouse PD-L1 antibody | BioLegend | Cat # 124307; RRID: AB_2073557 |
| PE mouse isotype antibody | BioLegend | Cat # 400607; RRID: N/A |
| APC mouse isotype antibody | BioLegend | Cat # 400611; RRID: N/A |
| Pacific Blue PD-1 antibody | BioLegend | Cat # 329915; RRID: AB_1877194 |
| PE-Cy7 PD-L1 antibody | BioLegend | Cat # 329717; RRID: AB_2561686 |
| Pacific Blue isotype antibody | BioLegend | Cat # 400151; RRID: N/A |
| PE-Cy7 isotype antibody | BioLegend | Cat # 400302; RRID: N/A |
| TruStain FcX | BioLegend | Cat # 422301; RRID: N/A |
| PE PD-1 antibody | Thermo Fisher Scientific | Cat # 12-9969-41; RRID: AB_10733013 |
| PE PD-L1 antibody | Thermo Fisher Scientific | Cat # 12-5983-41; RRID: AB_11042721 |
| BV421 CD45 antibody | BioLegend | Cat # 368521; RRID: AB_2687374 |
| Alexa700 CD11b antibody | BioLegend | Cat # 101222; RRID: AB_493705 |
| PE/Cy5 CD11c antibody | BioLegend | Cat # 301609; RRID: AB_493579 |
| Alexa488 CD3 antibody | BioLegend | Cat # 300319; RRID: AB_493690 |
| PE PD-L1 antibody | BioLegend | Cat # 329705; RRID: AB_940366 |
| APC PD-1 antibody | BioLegend | Cat # 329907; RRID: AB_940473 |
| PE isotype antibody | BioLegend | Cat # 400313; RRID: N/A |
| APC isotype antibody | BioLegend | Cat # 400119; RRID: N/A |
| AF647 mouse TCRβ antibody | BioLegend | Cat # 109217; RRID: AB_493347 |
| Mouse PD-1 blockade antibody (J43) | Thermo Fisher Scientific | Cat # 16-9985-82; RRID: AB_469307 |
| Isotype antibody for J43 | Thermo Fisher Scientific | Cat # 16-4888-81; RRID: AB_470171 |
| Human PD-1 blockade antibody (Pembrolizumab) | BioVison Inc | Cat # A1306; RRID: N/A |
| Isotype antibody for Pembrolizumab | BioLegend | Cat # 403701; RRID: N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) | Avanti Polar Lipids | Cat # 850457C |
| 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl) iminodiacetic acid) succinyl] (nickel salt, DGS-NTA-Ni) | Avanti Polar Lipids | Cat # 790404C |
| 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- (lissamine Rhodamine B sulfonyl) (ammonium salt, Rhodamine-PE) | Avanti Polar Lipids | Cat # 810158C |
| N-(4,4-Difluoro-5,7-Dimethyl-4-Bora-3a,4a-Diaza-s-Indacene-3-Propionyl)-1,2-Dihexadecanoyl-sn-Glycero-3-Phosphoethanolamine (Triethylammonium Salt, BODIPY-PE) | Thermo Fisher Scientific | Cat # D3800 |
| Poly-D-Lysine | Sigma-Aldrich | Cat # P6407 |
| CLIP-Surface 547 | New England Biolabs | Cat # S9233S |
| SNAP-Surface Alexa Fluor 647 | New England Biolabs | Cat # S9136S |
| SNAP-Cell 505-Star | New England Biolabs | Cat # S9103S |
| SNAP-Cell TMR-Star | New England Biolabs | Cat # S9105S |
| SNAP-Cell 647-SiR | New England Biolabs | Cat # S9102S |
| SEE super antigen | Toxin Technology | Cat # ET404 |
| Strep-SNAP-PD-1EX-His10 | This study | N/A |
| Strep-SNAP-PD-1EX | This study | N/A |
| Strep-SNAP-PD-1(K78A)EX -His10 | This study | N/A |
| Strep-SNAP-PD-L1EX-His10 | This study | N/A |
| Strep-SNAP-PD-L2EX-His10 | This study | N/A |
| Strep-SNAP-B7.2EX-His10 | This study | N/A |
| Mouse MHC-I H2Kb | Enfu Hui | N/A |
| Mouse PD-1EX-His | Sino Biological | Cat # 50124-m08h |
| Mouse PD-L1EX-His | Sino Biological | Cat # 50010-m08h |
| Mouse B7.2EX-His | Sino Biological | Cat # 50068-m08h |
| Mouse ICAMEX-His | Sino Biological | Cat # 50440-m08h |
| SIINFEKL peptide | Anaspec | Cat # AS-60193-1 |
| Live/Dead Aqua | Thermo Fisher Scientific | Cat # L34966 |
| Critical Commercial Assays | ||
| Human IL-2 ELISA MAX Deluxe | BioLegend | Cat # 431804 |
| CytoTox 96 Non-Radioactive Cytotoxicity Assay kit | Promega | Cat # G1780 |
| Cell Line Nucleofector Kit L | LONZA | Cat # VACA-1005 |
| Quantum R-PE MESF | Bangs Laboratories Inc | Cat # 827 |
| Quantum APC MESF | Bangs Laboratories Inc | Cat # 823 |
| Experimental Models: Cell Lines | ||
| HEK293T | Ronald Vale | N/A |
| Jurkat E6.1 T cells | Arthur Weiss | N/A |
| Raji B cells | Ronald Vale | N/A |
| HEK293F | Andrew Ward | N/A |
| Jurkat T cells with PD-1-mGFP | Enfu Hui | N/A |
| Raji B cells with PD-L1-mCherry | Enfu Hui | N/A |
| OT-I | Ananda Goldrath | N/A |
| EL4 | Ira Mellman | N/A |
| Oligonucleotides | ||
| See Table S2 for the list of Oligos | N/A | N/A |
| Recombinant DNA | ||
| See Table S3 for the list of recombinant DNA | N/A | N/A |
| Software and Algorithms | ||
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| Micro-Manager | Open Imaging | https://micro-manager.org/ |
| AccPbFRET | (Roszik et al., 2008) | http://biophys.med.unideb.hu/accpbfret/ |
| GraphPad Prism 5 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| FlowJo | FlowJo, LLC | https://www.flowjo.com/ |