Fig. 4. Analysis of tumor microenvironment in murine and human TNBC.

(A) Quantification of tumor infiltrating CD8⁺ T cells, represented as percentage of CD3⁺ cells (left) and per gram of tumor (right) in Py8119 TNBC tumors (n=6) treated with indicated mAbs (FACS analysis 22 days following tumor inoculation). (B-C) Representative images showing CD8⁺ T cell infiltration into Py8119 TNBC tumors (n=6) (B) and quantification of CD8⁺ cells as percentage of DAPI⁺ cells (C). (D-E) Contour plot showing migratory cross-presenting DCs, defined as CD45⁺/CD3⁻/F4/80⁻/CD11c⁺/MHC-II^high/CD103⁺/CD11b⁻ cells (D) and quantification of these cells as percentage of DCs (CD45⁺/CD3⁻/F4/80⁻/CD11c⁺/MHC-II^high) and total count (E). (F-G) Contour plot showing intra-tumoral F4/80⁺ macrophages, defined as CD45⁺/CD3⁻/Gr1⁻/CD11b⁺/MHC-II⁺/F4/80⁺ cells (F) and quantification of these cells as percentage of CD45⁺ CD3⁻ cells (top) and per gram of tumor (bottom) (G). (H) Human TNBC tumor sections stained with indicated markers. Serial sections were stained with DAPI and antibodies specific for CD8, E-cadherin and vimentin (panel #1) and DAPI, SOX4 and αSMA (panel #2). Data are summarized as mean ± SD. Data in [B, C] is an average of two independent experiments and data in [A, D, E, F and G] are representative of at least two independent experiments. For box plots, dots denote all individual values, horizontal lines denote median values, boxes extend from 25th - 75^th percentile of each group’s distribution, and no data points were excluded. An unpaired Student’s t-test was used to determine statistical significance, ***P < 0.001; **P < 0.01; *P < 0.05. See also Figure S4 and S5.