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Oltean et al. 10.1073/pnas.0603090103.

Supporting Information

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Supporting Figure 6
Supporting Table 1
Supporting Figure 7
Supporting Figure 8

Fig. 6. Fluorescence cytometry analysis of pRint and pRIIIcI² stably transfected DT3 cells (left two images) and AT3 cells (right two images). M1 represents a marker indicating fluorescence positive readings (i.e., above the background fluorescence for untransfected cells). The expression of red fluorescent protein (RFP) is relatively homogeneous, although it is clear that a small fraction of the Rint cells may have lost the reporter.

Fig. 7. Mesenchymal-epithelial transitions (MET) in tumors. (A) Zonula occludens protein-1 (ZO-1) counterstaining is consistent with MET. Example of a section from RIIIcI² tumors shows that the red fluorescent cells also stained positive for ZO-1. Images were acquired at ´200 magnification. (B) Vimentin counterstaining. Example of a section from RIIIcI² tumors shows that the red fluorescent cells have no or have low vimentin expression. The RFP+ cells are within a larger tumor region that has low vimentin staining. Images were acquired at ´200 magnification.

Fig. 8. A metastatic nodule does not express RFP, whereas in the lung tissue around it there are RFP-expressing metastatic cells. (A) Phase-contrast and hematoxylin/eosin staining of a large metastatic nodule in the lung of an animal bearing an AT3-RIIIcI² tumor (´40 magnification). (B) (Upper) Fluorescence pictures of lung tissue in the vicinity of the nodule, middle of the nodule, and at the interface of the nodule with the lung tissue. (Lower) Phase-contrast images of the same fields. Images were acquired at ´400 magnification.

*Frequency = No. of sections with at least one RFP+ island/no. of sections examined. Note that 12 of 14 tumors had at least one RFP+ cluster.

†Analysis of tumor sections with one RFP+ island that were costained with E-cadherin (see Fig. 3).

‡Stromal regions were identified by using Masson’s Trichrome stain (see Fig. 4).

§Analysis of lung sections with one RFP+ island that were costained with E-cadherin (see Fig. 5).

¶NA, not applicable; ND, not determined.

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