Effect of MAPK and Akt pathways in the denbinobin-induced COLO 205 cells apoptosis. A: Cells were pretreated with various kinds of MAPKs inhibitors; U10126 (U, 100 nmol/L), SB203580 (SB, 20 μmol/L), or SP600125 (SP, 100 nmol/L) for 1 h. The whole cell proteins were extracted from the cultured COLO 205 cells, which had been grown in 10% FCS and incubated for the indicated times with 0.1% DMSO or 20 μmol/L denbinobin in 0.1% DMSO for 24 h. After electrophoresis, proteins were transferred onto Immobilon-P membranes, and then probed with proper dilutions of specific antibodies; B: Effect of Akt pathway on the denbinobin treatment to COLO 205 cells. Time course study of 20 μmol/L denbinobin-treated COLO 205 cells was used to detect the total Akt proteins and the phorphorylated Akt by Western blot.