Published: Vol 4, Iss 9, May 5, 2014 DOI: 10.21769/BioProtoc.1120 Views: 10582
Reviewed by: Lin Fang
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Abstract
We sought to understand the mechanisms behind the potent effect of stromal TGF-beta program on the capacity of colorectal cancer (CRC) cells to initiate metastasis. We discovered that mice subcutaneous tumors and metastases generated in the context of a TGF-beta activated microenvironment displayed prominent accumulation of p-STAT3 in CRC cells compared with those derived from control cells. STAT3 signaling depended on GP130 as shown by strong reduction of epithelial p STAT3 levels upon GP130 shRNA-mediated knockdown in CRC cells.
Materials and Reagents
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Representative data
Figure 1. p-STAT3 staining of liver metastases generated after intrasplenic injection of CRC cells. Note strong staining in epithelial cells (arrows). E: epithelial cells, Str: stromal cells. Scale bar = 50 µm
Recipes
Acknowledgments
A.C. and D.V.F.T hold a Juan de la Cierva postdoctoral fellowship, E.E. an FPI PhD fellowship (both from Spanish Ministry of Science and Innovation). This work has been supported by grants from Instituto de Salud Carlos III FEDER (RD09/0076/00036) and the “Xarxa de Bancs de tumors sponsored by Pla Director d’Oncologia de Catalunya (XBTC), to E.B. from the European Research Council (Starting grant - 208488) and Consolider programmes (MICINN), to E.S. and A.C. by the Spanish Ministry of Science and Innovation, to JM by NIH grant CA34610 and to RM by grants PS09/00965 (MICINN) and NanoCoMets (CIBERBBN).
References
Article Information
Copyright
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Calon, A., Espinet, E., Palomo-Ponce, S., Tauriello, D. V. F., Iglesias, M., Céspedes, M. V., Sevillano, M., Nadal, C., Jung, P., Zhang, X. H., Byrom, D., Riera, A., Rossell, D., Mangues, R., Massague, J., Sancho, E. and Batlle, E. (2014). Immunostaining Protocol: P-Stat3 (Xenograft and Mice). Bio-protocol 4(9): e1120. DOI: 10.21769/BioProtoc.1120.
Category
Cancer Biology > General technique > Biochemical assays
Cell Biology > Tissue analysis > Tissue staining
Biochemistry > Protein > Immunodetection
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