Fig 5.

Association of the pri-miR-7 RNA with the QKI isoforms. (A) Schematic illustration of PCR primers used to detect the pri-miR-7-1 mRNA, the hnRNPK pre-mRNA, and mature mRNA. (B and C) HEK293 cells expressing pEGFP/hnRNPK (B) and pEGFP/hnRNPK:mQRE (C) were transfected with pcDNA or the indicated myc-QKI isoform. After 48 h, the cells were lysed and 10% of the fraction was used as input, while the remaining cellular extracts were immunoprecipitated (IP) with the anti-Myc antibody. The bound RNAs were purified and detected by semiquantitative RT-PCR to visualize the presence of the indicated RNAs. The samples were analyzed in the presence (+) or absence (−) of reverse transcriptase (RT) as indicated. The size of the DNA fragments is shown in base pairs on the right. (D) Cross-linked U343 cells were immunoprecipitated with either control IgG or anti-QKI-5 antibodies. The bound RNAs were isolated, and qRT-PCR was used to determine levels of bound pri-miR-7-1 or control mRNAs for GAPDH and HPRT.