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. 2016 Jul 30;7(35):56598–56607. doi: 10.18632/oncotarget.10967

Figure 4. Activated-Notch1 binds to RBP-Jκ binding site in CD133 promoter.

Figure 4

(A) Computer analysis revealed a RBP-Jκ binding site in CD133 promoter region. The putative binding site is indicated in bold. (B) KATOIII cells were transfected with either empty vector or pN1ICD-V5. Cross-linked chromatin was immunoprecipitated with anti-V5 antibody and analyzed by PCR using primers that flank the RBP-Jκ binding site. (C) KATOIII cells were transfected with RBP-Jκ siRNA. (left panel) RBP-Jκ expression was assessed by RT-PCR. (right panel) CD133 expression was evaluated by western blot. Densitometric analysis of RBP-Jκ and CD133 was performed. (D) AGS cells were transfected with either control siRNA or RBP-Jκ siRNA followed by either the control empty vector or N1ICD expressing plasmid transfection. CD133 and G3PDH expression was evaluated by RT-PCR. All of these experiments were performed thrice. The results are shown as means ± S.E. *P < 0.05 compared to control cells.