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. Author manuscript; available in PMC: 2011 Dec 1.
Published in final edited form as: Surgery. 2010 Dec;148(6):1228–1236. doi: 10.1016/j.surg.2010.09.026

Figure 3. Withaferin A inhibits RET protooncogene in human medullary thyroid cancer cells in vitro.

Figure 3

Human medullary thyroid cancer TT cells and DRO 81-1 cells were treated with increasing concentration of withaferin A for 24 hr and activity (measured as RET-phosphorylation) and total RET proto-oncogenes levels were analyzed using specific antibodies. Membranes were stripped and reprobed with anti-beta actin antibody to ensure equal protein loading and quantitative densiometry is shown below to compare protein expression as a normalized ratio to beta-actin levels. Withaferin A strongly inhibited Tyr905 phosphorylation of Ret at 3.0 uM concentration in TT cells and it reduced expression of total RET protein expression. In DRO 81-1 cells, both phospho-RET and total RET expression are inhibited at 5μM WA concentrations. Both cell lines demonstrate that Withaferin A is functional RET inhibitor in MTC cells.