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. 2015 Feb 24;25(4):459–476. doi: 10.1038/cr.2015.24

Figure 5.

Figure 5

The RNA-binding property of ADAR1 affects miR302 processing from pri-miR302s. (A) Rescue of ADAR1 expression in H9 cells by Flag-ADAR1, Flag-E912A or Flag-EAA. Left, a drawing showing the generation of ADAR1-rescued ADAR1 KD H9 cells by lentiviral infection. Right, WB confirms their expression in ADAR1 KD H9 cells. (B) Immunoprecipitation of Flag-ADAR1, Flag-E912A (catalytically inactive mutant), or Flag-EAA (RNA-binding-null mutant) from HEK293 cells extracts, followed by immunoblotting with ADAR1, DGC8 and DICER antibodies. (C) Flag-E912A, but not Flag-EAA, reversed the upregulation of miR302s in ADAR1 KD H9 cells. RT-qPCR revealed the expression of miR302s, miR30a, miR30e and miR577 (normalized to U6) in control, ADAR1 KD, KD+Flag-ADAR1, KD+Flag-E912A and KD+Flag-EAA H9 lines. Error bars represent ± SD of triplicate experiments. P values from one-tail t-tests are shown. Also see Supplementary information, Figure S5.