Fig. 7.

5A6 inhibits metastasis of the epitope knockin mouse breast cancer tumor 4T1-luc in a syngeneic mouse. (A) Flow cytometry analysis of Eat2 and 5A6 binding on parental 4T1-luc or 4T1-luc^5A6KI, the 5A6 epitope KI cell line. (B) 4T1-luc^5A6KI cells were placed at the top chamber of a Transwell assay and incubated with 5A6 or isotype control mAb; migrating cells attached at the bottom well were stained with crystal violet and quantified. Shown are mean ± SEM, Student’s t test, P = 0.0001***. (C) Schema: Mice were injected intravenously with 2.5 × 10⁴ 4T1-luc^5A6KI, followed by treatment on days +1, +3, and +5 with 100 µg of 5A6 MsIgG2a or an isotype control. Full body images were analyzed on day 16, then mice were killed, and lung metastases were analyzed ex vivo by (D) bioluminescence (IVIS imaging). Median is shown. Student’s t test, P = 0.0035** (two independent experiments, n = 15 in each group).