Journal: Cancers

Article Title: BCL2L11 Induction Mediates Sensitivity to Src and MEK1/2 Inhibition in Thyroid Cancer

doi: 10.3390/cancers15020378

Figure Lengend Snippet: Constitutively active AKT increases resistance to combined dasatinib and trametinib. ( A ) 8505C empty vector or Myr AKT cells were treated with indicated concentrations of dasatinib, trametinib, or the combination for 24 h and analyzed by Western blot for confirmation of pAKT. ( B ) CellTiter Glo assay for viability was performed on 8505C cells transfected with empty vector or myristoylated AKT (Myr AKT). Cells were treated with indicated doses of dasatinib and with 100 nM trametinib for 72 h. Area under the curve was calculated, and the results shown are mean ± SEM 2-way ANOVA * p < 0.05 ** p < 0.005. ( C ) Cleaved caspase 3/7 activity was measured after parental, empty vector, and Myr AKT cells were treated with DMSO, 100 nM trametinib, 50 nM dasatinib, or the combination for 24 h. Results shown are mean ± SEM. 2-way ANOVA test **** p < 0.003 * p < 0.03 ( D ) 8505C empty vector or Myr AKT cells were treated with indicated concentrations of dasatinib, trametinib, or the combination for 24 h and analyzed by Western blot for expression of BIM. Alpha tubulin was used as a loading control. Numbers below represent densitometric analysis normalized to loading control, followed by DMSO-treated cells. The uncropped blots are shown in .

Article Snippet: Antibodies were purchased from Cell Signaling: BIM (2933), Beta Actin (3700), pAKT S473 (4060), Total AKT (2920), pSrc Y416 (6943), Total Src (2109), BD Biosciences: Total FAK (610087), Invitrogen: pFAK Y861 (44-626G), and CalbioChem: α-tubulin (CP06).

Techniques: Plasmid Preparation, Western Blot, Glo Assay, Transfection, Activity Assay, Expressing, Control

Journal: Cell Death & Disease

Article Title: Targeting Btk/Etk of prostate cancer cells by a novel dual inhibitor

doi: 10.1038/cddis.2014.343

Figure Lengend Snippet: Inhibition of cell signaling in PC3 cells following treatment with CTN06. Cells were grown in 100-mm plate to 50% confluence and treated with CTN06 at the indicated concentrations. Cells were harvested after 12 h. pEtk, Etk, pBtk, Btk, pSrc, Src, pPLC γ 2, PLC γ 2, pStat3, Stat3, pAkt, Akt, pERK and ERK levels were measured using the corresponding antibodies by western blot. One of three similar experiments depicted

Article Snippet: Cells were grown in 100 mm dishes to about 50% confluence and treated with vehicle (control) and CTN06 (0.5 and 2 μ M) for 12 h. Proteins were detected by the following antibodies: β -actin (Sigma-Aldrich, A1978), Btk (Santa Cruz Inc., sc-1107), pBtk (pY233, Abcam, ab68217), Etk (Santa Cruz Inc., sc8874), pEtk (pY40, 3211S), Src (2109), pSrc (pY416, 6943), PLC γ 2 (3872), pPLC γ 2 (pY759, 6943), Stat3 (12640), pStat3 (pY705, 4113), ERK (4696) and pERK (4695) were obtained from Cell Signaling Inc. (Beverly, MA, USA).

Techniques: Inhibition, Western Blot