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

FIGURE 2.

FIGURE 2.

Effect of placing the HP in the 3′ UTR on Csy4-mediated knockdown. (A) Schematics of the 3′ UTR-HP reporter (left) and 3′ UTR-HP-Δp(A) reporter (right). (B) PCR products of randomly primed RNA isolated from HEK293 cells transfected with the GFP-3′ UTR-HP reporter (left) or GFP-3′ UTR-HP-Δp(A) reporter (right) and Csy4 as indicated. (C) Fluorescent images of HEK293 cells expressing GFP-3′ UTR-HP (left) or GFP-3′ UTR-HP-Δp(A) (right) in the absence (−) or presence (+) of Csy4. Corresponding transmitted light images are shown as insets in each fluorescent image. (D) Quantitation of Gluc activity by luminometric analysis at 24 h post-transfection for GLuc-3′ UTR-HP (left) or GLuc-3′ UTR-HP-Δp(A) (right) reporters. Error bars indicate standard deviation of four replicates. Statistical significance was calculated using a two-tailed Student's t-test ([****] P ≤ 0.0001). (E) PCR products of oligo-d(T)20 primed RNA isolated from HEK293 cells expressing GLuc-3′ UTR-HP (left) or Gluc-3′ UTR-HP-Δp(A) (right) with or without Csy4.