Figure 3.

HuR interacts with TM 5′UTR. (A) In vitro transcribed MS2hp and MS2hp-TM 5′UTR RNAs were incubated with cell lysates of A549 cells. After 1 h, the recombinant GST-MS2 proteins were added to the reaction mixture to perform a GST pulldown assay. The pulldown complexes were analyzed by SDS-PAGE and detected by HuR antibodies. (B) HuR binds with TM 5′UTR in A549 cells. Cells were transfected with pGL3 vector and pTM 5′UTR vector, and the cytosol extracts were immunoprecipitated with anti-HuR antibodies. The RNA associated with immunoprecipitated complex was extracted by Trizol reagent, and the level of HuR bound firefly luciferase mRNA (HuR-Luc) was detected by RT-PCR. The mRNA expression levels of Luciferase and GAPDH and the protein expression levels of HuR and tubulin were shown as an internal control.