Fig 10.
Nucleolin associates with ARF mRNA and mediates the p19ARF induction in HuR knockdown cells. (A) Lysates prepared from MEFs expressing sh-SCR or sh-HuR were immunoprecipitated using control (IgG) or nucleolin antibodies. RNAs were recovered from the immune complexes and analyzed by real-time PCR. (B) NIH 3T3 cells expressing sh-SCR or sh-HuR were transiently transfected with ARF (Δ5′UTR or full length)-expressing plasmids together with Flag-tagged nucleolin-expressing plasmids. Two days later, lysates were prepared and immunoprecipitated using control or Flag tag (M2) antibodies. RNAs in the immune complexes were analyzed by real-time PCR. Error bars represents SEM of results from triplicate samples. (C) MEFs (P2) expressing sh-SCR or sh-HuR were transfected with siRNA targeting nucleolin. Two days later, lysates were prepared and the expression of the indicated proteins was analyzed by immunoblotting. Lamin A/C was used as a loading control.