Figure 6.
The binding of nucleolin protein with EGF and PDGF‐BB mRNA in VSMCs. A, Binding of nucleolin protein to PDGF‐BB mRNA; B, binding of nucleolin protein to EGF mRNA Immunoprecipitation and RT‐PCR were used to analyse the combination of nucleolin and EGF, PDGF‐BB mRNA. Normal VSMCs and Ang II‐treated VSMCs were collected to prepare the cell extracts, and cell extracts were divided into three equal groups as follows: Input group, negative control IgG group and nucleolin antibody group. The nucleolin antibody was used for co‐immunoprecipitation, and the total RNA in half of the precipitate was extracted. The EGF, PDGF‐BB and β‐actin specific primers were used for RT‐qPCR detection, and the other half of the precipitate was detected by Western blotting analysis for the amount of nucleolin. Representatives of three separate experiments. RT‐qPCR, reverse transcription‐quantitative polymerase chain reaction; Ang II, angiotensin II treatment (10−5 mmol/L Ang II for 48 h); Input, positive control; IgG, immunoglobulin G negative control; Nuc‐Ab, nucleolin antibody; Ctrl, control cells; IP, immunoprecipitation