Menin represses malignant phenotypes of melanoma through regulating multiple pathways
Article first published online: 24 OCT 2011
© 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd
Journal of Cellular and Molecular Medicine
Volume 15, Issue 11, pages 2353–2363, November 2011
How to Cite
Gao, S.-B., Feng, Z.-J., Xu, B., Chen, Y., Zheng, H.-H., Yin, P., Hua, X. and Jin, G.-H. (2011), Menin represses malignant phenotypes of melanoma through regulating multiple pathways. Journal of Cellular and Molecular Medicine, 15: 2353–2363. doi: 10.1111/j.1582-4934.2010.01222.x
- Issue published online: 24 OCT 2011
- Article first published online: 24 OCT 2011
- Accepted manuscript online: 3 DEC 2010 10:41AM EST
- Received: August 18, 2010; Accepted: November 24, 2010
Table S1 Primer sequences and target sequences of shRNA
Table S2 Antibody and reagent
Table S3 Summarize of IHC results from certain primary melanoma samples
Fig. S1 (a) The proliferation of B16 with MEN1 knockdown. (b) The proliferation of A375 cells stably transfected with either empty vector or menin. (c) Migrated to lower side of the filter A375 cells were stained with 0.1% crystal violet. (d) Stably transfected A375 cells were added to the upper filter, and cell migration was determined, *P < 0.05, N = 3.
Fig. S2 (a) IF detection of menin (green), pFAK (green), DAPI (blue) and merge in the A375 cells. (b) PAK1-PBD agarose and Rhotekin RBD agarose were used to isolate GTP-Cdc42, GTP-Rac1 and GTP-RhoA from whole cell lysates from menin-overexpressing A375 cells. The Cdc42-GTP, Rac1-GTP and RhoA-GTP were detected using Western blotting and normalized by the total input protein. The pβ-catenin protein level was detected by Western blot in menin-overexpressing A375 cells.
Fig. S3 (a, c) Melanoma cells were treated with 1 μg/ml cisplatin or 250 μg/ml dacarbazine and harvested at various time-points. And the menin expression was determined with Western blotting. (b, d) Melanoma cells were treated with the indicated concentrations of cisplatin or dacarbazine, and the menin expression was detected by Western blotting. (e) A375 cells were treated for 24 hrs with various doses of Cisplatin and then analysed for apoptosis via Annexin V-PI staining. (f) menin, γ-H2A.X, cyclinB1 and cyclinB2 protein level were detected by Western blot.
Fig. S4 Primers for determining whether menin mutated were used.
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