Human miRNA Precursors with Box H/ACA snoRNA Features
Figure 5
snoRNA-like miRNA precursors that bind dyskerin.
Nuclear extracts were prepared from HeLa cells stably expressing either free GFP or YFP-dyskerin (YFP-DKC) and immunoprecipitated using an anti-GFP antibody. A Western blot confirming specificity of the immunoprecipitation using an anti GFP antibody. The same membrane was reprobed with an antibody against lamin as a loading control. B Position of the primers used to detect the specified miRNA extended regions. C RT-PCR used to detect co-precipitated hsa-mir-664, hsa-mir-151, hsa-mir-605, has-mir-215 and has-mir-140 miRNA precursors, with E2 box H/ACA snoRNA as positive control and hsa-pri-let-7g miRNA, U3 box C/D snoRNA, U1 snRNA, 5S rRNA and GAPDH pre-mRNA as negative controls for dyskerin-associated RNAs. The lane numbering in panel C refers to the lanes shown in panel A.