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. Author manuscript; available in PMC: 2021 Feb 24.
Published in final edited form as: Nat Cell Biol. 2020 Aug 24;22(9):1130–1142. doi: 10.1038/s41556-020-0560-6

Fig. 8. Fbxl7/FBXL7 suppression promotes pancreatic cancer metastasis, which is counteracted by decitabine or dasatinib treatment.

Fig. 8.

a, 1×106 FC1242 cells (parental or Fbxl7 KO) were injected in the spleen of 8-weeks old C57BL/6 mice. Sixteen days after injection, livers were collected, weighed, and photographed. n = 11 mice/group. Scale bar: 1 cm.

b-c, 80,000 FC1242 cells (parental or Fbxl7 KO) were injected in the pancreas of 8-weeks old C57BL/6 mice. Where indicated, two days after injection, dasatinib was administered by daily oral gavage (200 μl, 20 mg/kg). Eighteen days after injection, primary tumors (b), kidney, peritoneum, liver, epidydimis, and bladder were collected. In b, scale bar: 1 cm. n = 6, Parental-Untreated; n = 8, Parental-Dasatinib; n = 7, KO-Untreated; n = 6, KO-Dasatinib. In c, representative pictures of peritoneum, liver, and kidney with macrometastasis (arrows) are shown. Two independent experiments were performed. The table shows the number of mice with macrometastases/total number of mice. n = 17, Parental; n = 8, Parental + dasatinib; n = 18, KO; n = 6, KO+Dasatinib.

d, 500,000 AsPC1 cells stably expressing GFP-tagged NT or FBXL7 shRNAs were injected in the pancreas of 8-weeks old B6.129S7-Rag1tm1Mom/J(002216) mice. Five weeks after injection, primary tumors (top panels) and livers (bottom panels) were collected. Two independent experiments were performed. Scale bar: 1 cm. Top right panel, quantification of tumor weight (n = 15 mice/group). Bottom panels, livers were subjected to FACS analysis for GFP-positive cells (NT siRNA: n = 5 mice; FBXL7 siRNA: n = 4 mice).

e, 1×106 AsPC1 cells stably expressing GFP-tagged FBXL7 or NT shRNA were injected in the spleen of 8-weeks old B6.129S7-Rag1tm1Mom/J(002216) mice. Twenty-four hours before cell injection, decitabine was administered by intraperitoneal injection at 1 μg/g body weight. Six days after cell injection, livers were collected and subjected to FACS analysis for GFP-positive cells (NT siRNA: n = 5 mice; NT siRNA+ Decit.: n = 4 mice; FBXL7 siRNA: n = 5 mice; FBXL7 siRNA+Decit.: n = 6 mice). Two independent experiments were performed. Control: liver from normal, non-injected mouse. Decit.=decitabine. Mean ± s.d. is shown. P values are from unpaired, two-tailed t-test.