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. 2015 Jul 13;112(30):9418–9423. doi: 10.1073/pnas.1507087112

Fig. 3.

Fig. 3.

Anti-NCL 4LB5 scFV inhibits microRNA biogenesis. (A) HeLa cells, transfected as indicated, were incubated for 24 h with or without 120 nM 4LB5. The interaction between myc-tagged NCL and FLAG-tagged DGCR8 was assessed by coimmunoprecipitation followed by SDS/PAGE and Western blot. Control IgG was used as negative control to evaluate the specificity of the anti-Myc antibody for the immunoprecipitation. (B) REMSA performed by incubating recombinant NCL and biotin-labeled miR-21 in the presence of increasing amounts (80–650 nM) of 4LB5. Control IgG was used as negative control (CTRL). (C and D) NCL-dependent microRNA levels were analyzed by real-time PCR 72 h following 15 nM 4LB5 treatment. Both mature (C) and primary (D) miRNA levels were analyzed. Data are the average of three independent experiments performed in triplicate. *P < 0.05, **P < 0.01.