Fig. 5

CBP is a potential direct target of NLK in suppression of the NF-κB- and CREB-mediated transcription in LNCaP cells. a 48 h after transfection, Western blot analysis was used to detect protein levels of p65, IκBα, CREB and phosphorylated CREB (Ser133). For analysis of the subcellular distribution of p65, LNCaP cells were fractionated into cytoplasmic and nuclear fractions. b NLK and CBP are co-localized in the LNCaP cells. Cells were subjected to immunofluorescence with anti-NLK antibody followed by FITC-conjugated antibody (green) and anti-CBP antibody followed by Cy3-conjugated antibody (red). c Interaction of NLK with CBP in LNCaP whole cell extract. LNCaP cells were lysed and used for Co-IP assays with anti-NLK, anti-CBP or normal IgG antibody. d ChIP analysis of CBP occupancy of the Nurr1 promoter in LNCaP cells transfected with shRNA-control or shRNA-NLK. e 48 h after transfection, Western blot analysis was used to detect protein expression levels of CBP, NLK and Nurr1. f: 48 h after transfection, real-time PCR was performed to detect mRNA levels of Nurr1. Data are means ± SD for three independent experiments. *p < 0.05, assessed by Student’s t-test