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. Author manuscript; available in PMC: 2020 Jun 1.
Published in final edited form as: Prostate. 2019 Mar 22;79(8):896–908. doi: 10.1002/pros.23796

FIGURE 6. Exogenous palmitic acid promotes Src mediated oncogenic signaling and localization at the cell membrane.

FIGURE 6.

(A-B) Exogenous palmitic acid (PA) increases Src levels at the cell membrane fraction. SYF1 cells expressing Src(WT) were cultured in DMEM with 2% fatty acid free BSA. Cells were treated with or without 400 μM PA for 24 hours. Expression levels of Src kinase in the cytosol (Cyt) and the cell membrane (TM) fraction. Caveolin-1 and GAPDH were used as markers for TM and Cyt, respectively (A). Additionally, expression levels of Src kinase in the detergent soluble (S) and insoluble (I) membrane fractions were determined by immunoblotting. Tubulin and caveolin-1 were used as markers for detergent soluble (S) and insoluble (I) fractions, respectively (B). (C) 293T cells expressing doxycycline-inducible Src(Y529F) (293T+TRE/Src(Y529F) were grown in DMEM with 2% fatty acid free BSA. After treatment with dasatinib (10 nM) for 1 hour, the cells were further treated with DMSO, 200, or 400 μM of PA with/without doxycycline (1 μg/mL) for 24 hours. (D) PC-3 cells were grown in F-12K medium with 2% fatty acid free BSA and treated with DMSO, 300, or 600 μM of PA or PA and dasatinib (10 nm) for 24 hours. The levels of total Src, pSrc(Y416), total Fak, pFak(Y925), total Erk, and pErk1/2 were analyzed by Western blot. The data represent three independent experiments.