Skip to main content
. 2016 Dec 7;45(7):4036–4050. doi: 10.1093/nar/gkw1203

Figure 2.

Figure 2.

Knockdown activity and processing of AgoshRT5 variants. (A) The knockdown activity of the different AgoshRNA variants was determined by co-transfection with a luciferase reporter containing either the sense or antisense target sequence (upper and lower panel, respectively). The AgoshRNAs varied in the bottom bp. HEK293T cells were co-transfected with 100 ng of the respective firefly luciferase reporter plasmid, 1 ng renilla luciferase plasmid as internal control and 5 ng of the corresponding AgoshRNA construct. The regular shRT5 (21/5) was used as control. An unrelated shRNA (shNef) served as negative control, this activity was set at 100% luciferase expression. We performed three independent transfections, each in duplicate, and standard deviations were calculated. (B) Processing of the 3΄ strand (upper panel) and 5΄ strand (lower panel) of the AgoshRT5 variants was analyzed by northern blot. HEK293T cells were transfected with 5 μg of the indicated constructs. Size markers were included and length indicated in nt. The regular shRT5 (21/5) was included as control. An unrelated shRNA (shNef) was included as negative control. The regular shRNA ∼21 nt products are marked and * indicates the AgoshRNA ∼30 nt products. Ethidium bromide staining of small rRNAs and tRNAs are shown as loading controls below the blot.