Figure 7.

Monitoring E-cadherin Dynamics in Secondary Metastases from the E-cadherin-GFP Mouse

(A and B) Whole-body image of primary tumor (A) (B, white arrows) and isolated liver micro-metastases of a Kras^G12D/+; p53^R172H; E-cadherin-GFP mouse (B, red arrow). Blue arrow, auto-fluorescence in the bile duct.

(C–E) Western blot (C, top band, E-cadherin-GFP; bottom band, endogenous E-cadherin) and quantification (D and E) in isolated the primary and metastatic cell lines 101912 and 111375 (D) and 105925 (E).

(F and G) The isolated metastatic cell lines 101912 met (F) and 105925 met (G) from the liver of the E-cadherin-GFP mouse invading in organotypic matrices ± dasatinib (13 days, 200 nM). Black arrows, cluster formation upon dasatinib treatment. Shown is the quantification of cell invasion between 200–400 μm and cell clustering within matrices ± dasatinib.

(H and I) Immunofluorescence staining of E-cadherin within matrices ± dasatinib for the metastatic lines 101912 met (H) and 105925 met (I). Red arrows, junctions within cell clusters upon dasatinib treatment. E-cadherin, green; DAPI, blue.

(J and K) TEER (left) and Dispase assays (right) in metastatic lines derived from transgenic mouse, 101912 met (J), and 105925 met (K) ± dasatinib.

Columns, mean; bars, ± SE; n = 4 mice/group. ^∗p < 0.05 (unpaired Student’s t test). See also Figures S5 and S6.