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Figure 5 | Journal of Biomedical Science

Figure 5

From: Phospholipase A/Acyltransferase enzyme activity of H-rev107 inhibits the H-RAS signaling pathway

Figure 5

AACOCF3 and MAFP eliminated H-rev107 inhibition of H-RAS activation induced by EGF. (A, C) HtTA cells plated in 10-cm dishes were transfected for 24 h with H-rev107 or control vector in the presence of 10 μM AACOCF3, 5 μM MAFP, 2 μM BEL, 1 μM palmostatin B or DMSO vehicle. Cells were then serum starved for 12 h and stimulated with EGF (50 ng/mL) for 5 min. Cell lysates were prepared, and activated RAS (RAS-GTP) was collected with RAF-RBD-conjugated agarose. The levels of total RAS, H-rev107 fusion proteins, and actin were measured by Western blotting. (B, D) HtTA cells were plated in triplicate, incubated overnight, and then transfected for 2 days with control vector, H-rev107 expression vector, and reporter plasmids in the presence of 10 μM AACOCF3, 1 μM palmostatin B, 5 μM MAFP, 2 μM BEL, or DMSO vehicle. Cells were starved and then stimulated with EGF (50 ng/mL) for 12 h at 37°C. Cell lysates were prepared and analyzed for the transactivation activity of ELK1 protein. Representative results from triplicate samples are expressed as mean ± SD. Student’s t-test: **, P < 0.01; ***, P < 0.001.

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