Table 1.
Flow cytometry antibody cocktails used for the different endpoints
| Tube | FITC | PE (PacBlue for NK surface and biopsy) | PE-TR | APC | PerCP-Cy5.5 | APC-Cy7 |
|---|---|---|---|---|---|---|
| Biopsy | CD31 | Live/dead3 | CD563 | NKG2D4 | CD161 | CD451 |
| Biopsy isotype control | IgG11 | Live/dead3 | IgG23 | IgG11 | IgG14 | CD451 |
| EC | MICA/B2 | ULBP36 | CD563 | CD31 | CXCR34 | CD451 |
| EC isotype control | IgG22 | IgG26 | IgG23 | IgG11 | IgG14 | CD451 |
| NK cell surface marker | CD165 | NKp464 | CD563 | NKG2D4 | CXCR34 | CD451 |
| NK surface isotype control | IgG11 | IgG11 | IgG23 | IgG11 | IgG14 | CD451 |
| NK cell intracellular | Granzyme B*1 | IL-4*2 | CD563 | IFN-γ*2 | CD161 | CD451 |
| NK intracellular isotype control | IgG1*1 | IgG1*2 | IgG23 | IgG1*1 | IgG14 | CD451 |
CD45 was included in all cocktails (also in isotype control tubes) for differentiation between epithelial cells (ECs) and natural killer (NK) cells. PE, phytoerythrin; PacBlue, Pacific Blue; PE-TR, phytoerythrin-Texas red; APC, allophycocyanin; PerCP-Cy5.5, peridinin chlorophyll protein-cyanine 5.5; APC-Cy7, allophycocyanin-cyanine 7. The antibodies were purchased by the following companies: 1BD Biosciences, 2eBioscience, 3Invitrogen, 4Biolegend, 5Beckman Coulter/IO Test, and 6R&D.
*
These antibodies were added during the second staining step after fixation and permeabilization.