Figure 7.

RNA‐EMSA and Luciferase reporter gene assay of the binding of nucleolin to the 5′ UTR of EGF and PDGF‐BB mRNA. A, The binding of nucleolin to the 5′ UTR of EGF mRNA. B, The binding of nucleolin to the 5′ UTR of PDGF‐BB mRNA. Cyto extracts: cytoplasmic protein; pcDNA3.1: control plasmid; pcDNA3.1‐Nuc: overexpression plasmid; Nuc1‐309: nucleolin mutant with lacking the carboxy terminus of nucleolin (ie deleting the amino acid containing the RNA‐binding domain); anti‐nuc antibody: nucleolin antibody; biotin‐labelled probe: biotin‐labelled EGF 5′ UTR RNA probe or biotin‐labelled PDGF 5′ UTR RNA probe; 100‐fold competition probe: 100‐fold unlabelled probe; C, effect of Ang II on the binding activity of nucleolin and EGF mRNA; D, effect of Ang II on the binding activity of nucleolin and PDGF‐BB mRNA. Representatives of three separate experiments. Ang II: AngII treatment (10⁻⁵ mmol/L Ang II for 48 h); E, Luciferase reporter gene assay for the regulation of EGF mRNA 5′ UTR by nucleolin; F, Luciferase reporter gene assay for the regulation of nucleolin on PDGF‐BB mRNA 5′ UTR. Nuc1‐309: Nuc1‐309: nucleolin mutant with lacking the carboxy terminus of nucleolin (ie deleting the amino acid containing the RNA‐binding domain); Control: normal cell control group (ie no Ang II treatment group); Ang II: Ang II treatment (10⁻⁵ mmol/L Ang II for 48 h); pcDNA3.1‐Nuc: overexpression plasmid; pcDNA3.1: control plasmid; ^* P < .05 vs pcDNA3.1 and Nuc1‐309 group, ^# P < .05 vs pcDNA3.1 and Nuc1‐309 group