Figure 1. RAE-1δ is induced on tumor-associated macrophages in subcutaneously transferred and spontaneous tumors.
(A) Established B16 S.C. tumors were dissociated and analyzed for NKG2D ligand expression on tumor-associated macrophages. (B) RAE-1δ expression (left) and MFI quantification (right) on the indicated cell types in B16 tumors. (C) RAE-1δ expression on TAMs in spontaneous KP sarcoma, but not in spontaneous TRAMP prostate adenocarcinoma or transferred RMA-S lymphoma. Data are representative of >3 independent experiments.
Figure 1—figure supplement 1. Gating strategy and RAE1δ expression on tumor-associated macrophages and monocytes in mice with B16 tumors.
(A) Gating strategy to identify tumor-associated macrophages in B16 tumors. (B) Expression of RAE-1δ on TAMs in B16 S.C. tumors in WT and RAE-1-KO mice. (C) RAE-1δ expression on TAMs, splenic macrophages, blood monocytes, or peritoneal macrophages in WT mice with established B16 S.C. tumors. Data are representative of >3 independent experiments.
Figure 1—figure supplement 2. RAE-1 antibody validation and RAE-1δ staining on TAMs in different tumors. .
(A) B16 cells transduced to stably express RAE-1δ or RAE-1ε were stained with biotinylated antibodies against RAE-1δ or RAE-1ε, followed by fluorophore-conjugated streptavidin. (B) Expression of RAE-1δ on TAMs in B16 S.C. tumors in WT mice at the indicated time after injection of 1 × 106 tumor cells. (C) Expression of RAE-1δ on TAMs in S.C. B16 and RMA-S tumors and autochthonous KP and TRAMP tumors. Data are compiled from several independent experiments.
Figure 1—figure supplement 3. Gating strategies for blood and tumor-associated monocytes and peritoneal macrophages.
Gating strategies for (A) monocytes in B16 tumors, (B) blood monocytes, and (C) peritoneal macrophages.