FIGURE 1.
A3 mRNA expression profile in HeLa cells and verification of shRNA-mediated knockdown of A3 mRNAs. A, endogenous expression of A3 mRNAs in HeLA cells. mRNA copy numbers were determined by real-time RT-PCR and normalized to GAPDH. Shown is a representative experiment of at least three independent experiments. Values are mean ± S.D. of triplicates. B, efficiency of shRNA-mediated knockdown of A3 mRNAs. HeLa cells were transduced with lentiviruses carrying pSicoR-MS2 scrambled shRNA or shRNA against A3B or A3C mRNA. Successfully transduced cells were selected with puromycin for at least 1 week to obtain a pure polyclonal population. RNA was analyzed by real-time RT-PCR to determine the expression levels of A3B or A3C mRNA. Results were calculated by the ΔΔCT method using GAPDH as a control gene and normalized to cells infected with scrambled shRNA. Values are mean ± S.E. of at least three independent experiments.