Fig. 6

PKN2 inhibits the transcriptional activities of CREB and Elk-1. a HT-29cells were stably transfected with shCTL, shPKN2-1 or shPKN2-2 and treated with solvent or U0126 (1 μM) for 1 h. Western blotting was used to detect the indicated proteins. b Stable clones of SW480, HCT116 were detected for indicated proteins using western blotting. c RKO cells were transfected with PKN2-WT-HA, CREB-WT-flag, Elk-1-WT-flag or control vectors. IL4 and IL10 gene expression was detected. ***,P < 0.001 versus lane 1. ###, P < 0.001 versus lane 2. d Diagram shows the predicted binding sites of Elk-1 and CREB on the -1000 bp ~ + 1 bp promoter regions of IL4 and IL10. e ChIP assays of Elk-1 or CREB on the chemokine promoters in SW480 stably transduced with PKN2-WT or vector, and HT-29 cells transduced with shCTL or depletion of PKN2. These assays were repeated three times, and representative photos are shown