Figure 3. Gaining chromosome 5 induces a cell state transition in colon cancer cells.

(A) Western blot analysis of epithelial marker expression in HCT116 trisomies. Alpha-tubulin was used as a loading control.

(B) Western blot analysis for Fibronectin, Vimentin, and N-cadherin indicates that HCT116 trisomies do not express mesenchymal genes. The sarcoma cell line U2OS was analyzed as a positive control.

(C) Schematic of two strategies to select for HCT116 Ts5 chromosome-loss revertants.

(D) Western blot analysis of epithelial marker expression in trisomy 5 cells that lost the extra copy of chromosome 5. “Xeno”: cells isolated after xenograft growth. Ts5 c1 e3, e5, e10: clones isolated from high-passage cells.

(E-F) Quantification of the average number of cells per field that were able to cross the membrane in the invasion assay. Averages represent three independent trials in which 15–20 fields were counted.

(G-H) Verification of E-cadherin and EpCAM overexpression in two HCT116 Ts5 clones transfected with the indicated plasmids.

(I) Quantification of the average number of cells per field that were able to cross the membrane in the invasion assay. Averages represent three independent trials in which 15–20 fields were counted.

(J) Representative images of invasion in the indicated cell lines.

Bars represent mean ± SEM. Unpaired t-test *** p<0.0005. Scale bar, 50 μm