Fig. 6

Compounds D14 and C22 decrease the activation of ras in ondependent cells of KRas4B and the phosphorylation of AKT and ERK promoting the increased phosphorylation of p53. a Ras activation (Ras-GTP) decreases in the MIA PaCa-2 cell line treated with the compounds D14 (99.33 μM), C22 (137.5 μM) and Deltarasin at 5 μM for 3 h. b Representative immunoblot of whole protein extracts from MIA PaCa-2 treated with D14, C22 and deltarasin for 3 h, detected phosphorylation inhibition of AKT and ERK using GAPDH with load control. c and d The quantitative results of the immunoblot of 3 independent studies are shown in graphs. e D14 and C22 compounds induced inhibition of the phosphorylation of ERK1 (T202/Y204), ERK2 (T185/Y187) in human MIA PaCa-2 cancer cell and promoted an increased in the P53 phosphorylation (e and f). The phosphorylation of those proteins were significantly inhibited compared with the control. Protein extract (300 μg) were used for human phospho-MAPK array kit. Array spots were visualized in accordance with the manufactures’s instructions. The intensity of each spot was measured as described in “.Material and Methods”. The upper panel gives the array analysis from MIA PaCa-2 without compounds (control); the middle panel shows the array analysis of MIA PaCa-2 cell line treated with D14 compound; the lower panel shows the array analysis of MIA PaCa-2 cell line treated with the C22 compoud. e Normalized quantitation graph that shows the relative fold change of phosphorylation differences upont for control