Figure 5 from TGFβ Inhibition during Radiotherapy Enhances Immune Cell Infiltration and Decreases Metastases in Ewing Sarcoma

TGFβ inhibition during radiotherapy increases immune infiltration of Ewing sarcomas. A, Experimental schematic depicting the four treatment groups. B, Immunophenotyping was performed on single-cell suspensions derived from TC32 Ewing tumors established in hu-CD34+ mice. The total number of huCD45+ cells was determined in each tumor. Box plot represents average total hu-CD45+ count across three biologic replicates, line represents mean, and top and bottom limits of box are the top and bottom quartiles, respectively. *, P P > 0.05. C, Immunophenotyping was performed on huCD34+ tumor single-cell suspensions. Samples were stained using multispectral panel (see “Materials and Methods”). Samples were analyzed on a Cytek Aurora spectral flow cytometer and then unmixed and .fcs files were imported into FlowJo for manual gating. The manual gating strategy is available in Supplementary Fig. S15. Proportion of live, huCD45+ cells that are CD3+ or CD3− are shown (T cell marker). D, Proportion of CD3+ cells that are CD4+ T cells, regulatory T cells (Treg), or CD8+ cells are shown. E, Proportion of CD3− cells that are CD56+ (NK cell marker), CD14+ (monocyte marker), CD14+/CD11c+ (myeloid derived suppressor cell), and CD19/CD20+ (B cell marker) are shown. RT, radiotherapy. A, Created in BioRender. Daley, J. (2025) https://BioRender.com/jn2ii5x