Extended Data Fig. 11: EPOR expression on tumor-infiltrating leukocytes (TILs), tumor Ag-carrying migratory cDC1s in TDLNs and tumors, and the correlation of tumor growth with systemic EPO levels.
From: Erythropoietin receptor on cDC1s dictates immune tolerance
a, Zbtb46GFP/+EportdT/+ mice were implanted s.c. with MC38 or B16F10 tumor, or EO771 tumor in the mammary fat pad. On day 12, tumors were harvested for flow cytometric analysis of Epor-tdT on cDCs (live-dead aqua−CD45+Zbtb46-GFP+CD11c+); cDC1s were gated as XCR1+ and non-cDC1s as XCR1−. b-d, Mice implanted s.c. with MC38-OVAdim or B16F10-OVA; on day 10, tumors were analyzed for Epor-tdT expression in TILs. b, Representative gating strategy of individual live-dead blue− TIL populations. c,d, Histogram overlay showing Epor-tdT expression in individual cell populations. e, Zbtb46GFP/+EportdT/+ mice with MC38-OVAdim tumors (day 12) were analyzed for Epor-tdT on tumor-infiltrating cDCs; CCR7+ (population I) and CCR7− (populations II/III by Ly6A) subsets were gated, with XCR1/CD103 staining to define cDC1s and cDC2s. f, Quantification of Epor-tdT expression on individual tumor infiltrating cDC subsets. MC38-OVAdim (n = 4) and B16F10-OVA (n = 4) tumors were harvested on day 12 post-s.c. implantation for flow cytometry. Gating strategy as in Fig. 5a and Extended Data Fig. 11e. g, Flow cytometry analysis of Epor-tdT expression on TDLN migratory cDC1s. Overlay of migratory cDC1s with Lin− live cells to show Epor-tdT expression levels. h, Serum EPO levels were measured by ELISA on the indicated days after s.c. implantation of MC38-OVAdim (n = 6) or B16F10-OVA (n = 5) tumors in WT mice. i,j,k, B16F10-OVA-ZsGreen cells were s.c. implanted into EportdT/+ mice, and tdLN and tumor were analyzed on day 9 after inoculation. j, Flow cytometry analysis of Epor-tdT expression on tdLN ZsGreen+ migratory and resident XCR1+ cDC1s. Overlay of migratory ZsGreen+ cDC1s or resident ZsGreen+ cDC1s with Lin−live cells to show Epor-tdT expression levels. k, Flow cytometry analysis of Epor-tdT expression on tumor infiltrating ZsGreen+ cDC1s. Data are shown from one experiment, representative of at least two independent experiments with similar results (a-e,f,g,h,j,k). Statistical analysis was performed using one-way ANOVA with Dunnett’s multiple-comparison test (f). Data are mean ± s.e.m. (f,h). The diagrams in a,b,e,g,i were created in BioRender. Zhang, X. (2025) https://BioRender.com/gjjtedh.
