Figure 5
NF-κB pathway mediated DAB2IP-regulated EMT and stemness.
(A) Nuclear extracts were subjected to western blot analysis and p65 was determined. Histone H3 was used as nuclear marker and GAPDH was used as cytoplasmic marker. (B) luciferase reporter assay showed that NF-κB transcriptional activity was reduced in SW480-D2 compared with SW480-con when transiently transfected with scramble siRNA and was increased in SW48-D2 when transfected with si-IκB compared with SW480-D2-scramble. A strong increase of NF-κB transcriptional activity in HT29-KD6-scramble and HCT116-KD3-scramble was detected and it was reversed when transfected with si-p65. (C) EMT markers were analyzed by western blot in SW480-D2 when treated with scramble or si-IκB and in HT29-KD6 and HCT116-KD3 treated with scramble or si-p65. (D) SW480-D2 was transiently transfected with si-IκB and quantative analysis of spheres was done compared with SW480-con and SW480-D2 with scramble si-RNA. HT29-KD6 and HCT116-KD2 transfected with si-p65 were analyzed similarly.
