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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Nature. 2015 Nov 11;527(7579):472–476. doi: 10.1038/nature15748

Extended Data Figure 4. Characterization of Tri-PyMT cells.

Extended Data Figure 4

a, EMT of Tri-PyMT cells with TGF. RFP+ Tri-PyMT cells were sorted by flow cytometry and cultured in medium containing 2% FBS with or without TGF-β1 (2ng/mL) for 3 days. Plot shows quantification of the percentage of GFP+ cells analyzed by flow cytometry (n=2 biological replicates).

b, Cell migration assay of Tri-PyMT cells. The tracing plots show the movement of individual RFP+ and GFP+ cells in 10 hours of live imaging. Quantification plot (right panel) showed the average distance that RFP+ and GFP+ cells have moved during the time frame (n>20, *p<0.01).

c, Relative expression of epithelial, mesenchymal, and tumor markers in sorted RFP+ and GFP+ Tri-PyMT cells as determined by Q-RT-PCR with GAPDH as the internal control. n=2 individual experiments.

d, EMT of Tri-PyMT cells is reported by fluorescent marker switch. Flow cytometry plot shows E-cadherin- (E-Cad-) and E-cadherin+ (E-Cad+) subpopulations of Tri-PyMT cells (upper panel). Of the E-Cad- and E-Cad+ subsets, the populations were further dissected according to innate fluorescence (lower panel). Numbers indicate the percentage of GFP+, RFP+, or transitioning (Q2) cells in the parental E-Cad- or E-Cad+ subsets, respectively.