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Figure 1 | BMC Cancer

Figure 1

From: Selective inhibition of RET mediated cell proliferation in vitro by the kinase inhibitor SPP86

Figure 1

SPP86 selectively inhibits RET- induced ERK1/2 phosphorylation in thyroid cancer cell lines. (A) TPC1 cells were cultured overnight in media containing 0.1% FBS and exposed to the indicated concentrations of sorafenib or SPP86 for 90 min in similar media. Total lysates were resolved by SDS-PAGE and probed with antibodies directed against phosphorylated (Thr202/Tyr204) and total ERK1/2. Monoclonal antibodies directed against actin were used to monitor gel loading. (B) 8505C cells expressing mutant BRAFV600E and (C) C643 expressing mutant RASG13R were treated as in A. (D) TPC1 cells were treated as in C. Total lysates were resolved by SDS-PAGE and probed with antibodies directed against phosphorylated (Tyr1062), total RET, phosphorylated ERK1/2 (Thr202/Tyr204) and total ERK1/2. Monoclonal antibodies directed against tubulin were used to monitor gel loading. (E) TPC1 cells were cultured in media containing 0.1% FBS overnight and then exposed to the indicated concentrations of SPP86 for 20 h under the same conditions. Total lysates were resolved by SDS- PAGE and membranes were probed with the indicated antibodies. Monoclonal antibodies directed against PARP were used to monitor gel loading. (F) TPC1 cells cultured in media containing 0.1% FBS were exposed to the indicated concentration of SPP86 for 90 min. Total cell lysates were resolved by SDS- PAGE and probed with antibodies directed against phosphorylated and total RET or phosphorylated and total ERK1/2. PARP was used instead of actin or tubulin to monitor gel loading, to enable the simultaneous detection of Akt and ERK1/2 on the membrane. (G) 8505C and C643 cells were cultured overnight in media containing 0.1% FBS. Cells were exposed to the indicated concentrations of SPP86 for 20 h. Lysates were probed with antibodies directed against the indicated proteins.

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