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. 2015 Nov;21(11):1921–1930. doi: 10.1261/rna.051227.115

FIGURE 1.

FIGURE 1.

Csy4-mediated knockdown of 5′ UTR-hairpin (HP) and ATG-HP constructs. (A) Sequence of the Csy4 hairpin substrate and fragments after enzymatic cleavage. The cleavage site is indicated by a black triangle on the unprocessed hairpin. (B) Schematics of the unmodified control (left), 5′ UTR-HP (middle), and ATG-HP (right) reporters. (C) PCR products of randomly primed RNA isolated from HEK293 cells transfected as indicated. (Left) unmodified control, (middle) 5′ UTR-HP-GFP, and (right) ATG-HP-GFP. (D) Fluorescent images of HEK293 cells expressing unmodified control (left), 5′ UTR-HP (middle), and ATG-HP GFP (right) reporters in the absence (−) or presence (+) of Csy4. Corresponding transmitted light images are shown as insets in each fluorescent image. (E) Quantitation of GLuc activity by luminometric analysis at 24 h post-transfection for unmodified control (left), 5′ UTR-HP (middle), and ATG-HP (right) GLuc reporters. Error bars indicate standard deviation of four replicates. Statistical significance was calculated using a two-tailed Student's t-test ([****] P ≤ 0.0001).