Figure 4.

SAM, anti-PD-1 antibody, and the combination treatment decreased lung metastasis in Eo771 tumor-bearing mice. Briefly, mice were injected with Eo771 cells orthotopically at the m.f.p and treated with the four treatments indicated. At the end of the study, lungs of the mice were harvested, fixed using formalin, embedded in paraffin, sliced, and stained with H&E. (A) Representative histology images of mouse lung showing the whole lung and magnified images to show metastatic lesions from each group except the SAM+anti-PD-1 antibody combination group, which had no lesions in this sample. Lens: top; 4×; bottom; 20×. Magnification: top; 40×; bottom; 200×. (B) Total metastatic lesion area (µm² ± SEM) for each group (n = 4/group). Total metastatic lesion area was calculated by annotating all the metastatic lesions in the entire lung of a mouse using the ImageScope annotation tool, which gives the selected area. Then, all the lesion areas were added together. (C) Percentage of mice with lung metastasis in each group. Statistical significance was determined by (B) one-way ANOVA in GraphPad prism. Significance values are represented by asterisks (* p < 0.05; ** p < 0.01).