(A) Mammary glands were isolated from MMTV-PyMT at
different stages of tumor progression and tissue sections were stained with
H&E stain or immunostained with Slit2 antibody using the IHC technique.
B-E. (B) Eight weeks old MMTV-PyMT mice bearing spontaneous
mammary tumors were treated with rSlit2-N or PBS for four weeks.
(C) Tumor volume was measured every week up to 12 weeks of age.
(D) Representative pictures of the lungs harvested from (B).
(E) The number of metastatic nodules in the lungs.
F-M. MVT1 cells were orthotopically implanted into the six
weeks old FVB wild type mice. (F) Schematics of mice treatment with
rSlit2-N or PBS. N=6 mice in each group. (G) Tumor volume was
measured every week. (H) Representative pictures of the lungs
harvested from (F). (I) The number of metastatic nodules in the
lungs. (J) Schematics of mice treatment with rSlit2-N or PBS,
showing Slit2 treated tumors were allowed to grow for additional two weeks. N=5
mice in each group. (K) At the end, tumors were harvested and
weight was measured. (L) Representative pictures of the lungs
harvested from (J). (M) The number of metastatic nodules in the
lungs.
N-Q. (N) Human breast cancer cell line MDA-MB-231
overexpressing Slit2 (231-Slit2) or vector control (231-Vec) implanted
orthotopically into the NSG mice. (O) After two weeks of tumor
injection, tumor volume was measured every week up to the age of 12 weeks.
(P) The images of lungs harvested from (N). (Q)
The harvested lungs were analyzed for the number of metastatic nodules using a
dissection microscope.
R-U. NSG females were injected with MDA-MB-231 cells and
treated with Adeno-Slit2 or Adeno-Null. (R) Schematics of mice
treatment with Adeno-Slit2 or Adeno-Null. (S) Starting from 8 weeks
of age, tumor volume was measured every week. (T) Representative
pictures of the lungs harvested from (R). (U) The number of
metastatic nodules in the lungs. * is p<0.05, ** is p<0.01, *** is
p<0.001, NS is P value not significant using student’s t-test.