Journal: Cancer Research

Article Title: Mammary Tumor–Derived Transplants as Breast Cancer Models to Evaluate Tumor–Immune Interactions and Therapeutic Responses

doi: 10.1158/0008-5472.can-21-0253

Figure Lengend Snippet: Figure 3. Comparison of orthotopic and subcutaneous mTDT response to anti–PD-L1. A, Representative images of multiplex staining for CD8þ T cells (green), pSMAD2 (mauve), PD-L1 (red), F4–80 (yellow), pan-cytokeratin (white), and DAPI counterstained nuclei (blue) for parental tumors E, G, H, and I and corresponding orthotopic and subcutaneous mTDT. White scale bar, 20 mm. B, The percentage of stroma in orthotopic (orange circles) and subcutaneous (purple triangles) mTDT. C–F, The cell density (number per mm2) of CD8 (C), PD-L1 (D), F4–80 (E), and pSMAD2 (F) in mTDT from four infiltrated parents grown in orthotopic (n ¼ 5; orange circles) versus subcutaneous (n ¼ 4; purple triangle) sites. G, Tumor growth curves of mTDT from six infiltrated families treated with anti–PD-L1 (purple) versus control antibody (black dash). H, Overall survival curve for mTDT from six infiltrated families treated with control IgG (black dash; n ¼ 27) or anti–PD-L1 (purple; n ¼ 31). I, Combined Kaplan–Meier survival graph of mTDT from family B (red), family E (green), G (pink), H (orange), I (blue), and K (gold). Cox regression analysis table of factors affecting survival of mTDT shown as Kaplan–Meier. , P < 0.05; , P < 0.001.

Article Snippet: Slides were then incubated overnight at 4 Cwith primary antibodies as follows: CD3e monoclonal antibody (SP7; Thermo Fisher Scientific; cat. #MA5-14524, RRID: AB_10982026, 1:100), anti-CD8 (Abcam; cat. #ab4055, RRID:AB_304247, 1:100), CD11b (M1/70; BioLegend; cat. #101201, RRID: AB_312784, 1:100), purified anti-mouse CD45 (30-F11; BioLegend cat#103101, RRID: AB_312966, 1:100), pSMAD2 (Cell Signaling; cat. #3122S, RRID: AB_823638, 1:50), F480monoclonal antibody (SP115; Thermo Fisher Scientific; cat. #MA516363, RRID: AB_2537882, 1:100), anti-wide spectrum cytokeratin antibody (Abcam; cat#ab9377, RRID: AB_307222, 1:200) and recombinant anti–PD-L1 antibody (EPR20529; Abcam; cat. #ab213480, RRID: AB_2773715, 1:50).

Techniques: Comparison, Multiplex Assay, Staining, Control