Figure 2.

Gating strategy to examine immune cell landscape in tumor/lungs of tumor-bearing mice treated with PAMAM-G3 or PBS

The intravenous experimental lung metastasis 4T1 mouse model of breast cancer described in Figure 1 was used for this analysis. Lungs were collected from PAMAM-G3- or PBS-treated tumor-bearing mice on day 15 and processed for immune cell analysis via flow cytometry. Lungs from naive nontumor-bearing mice were used as controls. One representative mouse from each group is shown. (A) Hematopoietic cells in lungs were identified via CD45.2 cell surface expression. Immune cell populations were identified based on expression of CD11b (innate/myeloid), B220 (B cells), CD4 and CD8 T cells, and NK1.1 (NK cells). (B) Neutrophils within the CD45.2 gate were further identified as Ly6C⁺Ly6G⁺. Ly6C⁺Ly6Clo/neg cells that did not express Ly6G were analyzed for expression of Ly6C and CD64. Ly6C^hiCD64⁺ monocytes were examined for expression of MHC class II (MHC-II) and were identified as inflammatory monocytes (also known as monocyte-derived cells).