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. 2014 Mar 12;9(3):e91585. doi: 10.1371/journal.pone.0091585

Figure 7. Effect of neomycin and lincomycin on the FLAG-CRD-BP-mRNA interaction in cells.

Figure 7

HeLa cells were plated and transfected with pcDNA-FLAG-CRD-BP as described in Materials and Methods. Twenty-four hours after transfection, cells were treated with 750 μM of neomycin or lincomycin, or received no treatment. Immuno-precipitated FLAG-CRD-BP from cell lysate was subjected to RNA extraction and then quantitative real-time PCR (A) or Western blot analysis using anti-FLAG antibody as shown in (B). In a different set of experiments, HeLa cells transfected with pcDNA-FLAG-CRD-BP were directly subjected to total RNA extraction to assess steady-state levels of the c-myc, β-actin and CD44 mRNAs by quantitative real-time PCR (C). Total cell lysate was also analyzed to detect FLAG-CRD-BP protein expression in all groups, with β-actin used as a loading control (D). Data shown in (A) and (C) were pooled from three biological replicates.