Figure 10

TAR miRNA altered ERCC1 protein expression without altering mRNA levels. (A) 293T cells were transfected with psiCheckERCC-737 and either pPolIII TAR or pPolIIIScr. Renilla and firefly luciferase expression was measured after 72 hours. Data shown represent the normalized ratio of Renilla luciferase to firefly luciferase for two replicates. 293T cells were transfected with either pPolIII TAR or pPolIIIScr. Forty-eight hours after transfection the cells were harvested and protein and RNA extracts were prepared. (B) Protein extracts were Western blotted for ERCC1 and β-actin expression. (C) RNA extracts were used to generated cDNA, and ERCC1 and IER3 mRNA levels were determined by PCR.