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. 2020 Oct 23;18(8):1170–1180. doi: 10.1080/15476286.2020.1836457

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Figure 3. — Position dependence of pri-miRNA processing efficiency. (A) Processing efficiency was measured at multiple positions of the cluster. At each position, two pairs of PCR primers were used: for the total pri-miRNA level measurements, both primers are located outside the pre-miRNA stem-loop; for the unprocessed pri-miRNA levels, a forward primer on the pre-miRNA stem-loop and a reverse primer downstream from the pre-miRNA were used. (B) Processing efficiency shows a position-dependent difference between placenta (n = 4; blue dots) and ESC samples (n = 6, red dots) (β-coefficient: −0.014825, p value: 0.0184). (C) Processing efficiency difference between control (n = 2; blue dots) and ActD treated JAR cells (n = 2; purple dots) was not affected by chromosomal position (β-coefficient: 0.002638, p value: 0.9256). (D) Drosha depletion by siRNA (n = 2; orange dots) reduced processing efficiency in a position-dependent manner in HuES9 cells (β-coefficient: −0.0228201, p value: 0.00131). (E) In JAR cells, Drosha depletion (n = 2; purple dots) had no effect compared to the control siRNA treatment (n = 2, blue dots) (β-coefficient: −0.002625, p value: 0.881)