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. Author manuscript; available in PMC: 2010 Mar 10.
Published in final edited form as: Nature. 2009 Aug 23;461(7261):230–235. doi: 10.1038/nature08283

Figure 4. Effects of ds RNA produced by the hTERT-RMRP RdRP.

Figure 4

a, Semi-quantitative RT-PCR for total RMRP and retrovirally delivered RMRP (ectopic) in cell lines expressing control or RMRP expression vectors. Promoters used to express RMRP are indicated. The relative intensity of RMRP is noted below each panel. See Supplementary Fig. 14. b, RT-PCR for total RMRP. See Supplementary Fig. 15. c, Effects of suppressing hTERT on RMRP levels. A control shRNA (sh-GFP) or 2 different hTERT-specific shRNAs were stably introduced into HeLa cells. d, Effects of RMRP mutants on RMRP levels. RT-PCR was used to detect RMRP levels in (c, d). e, Detection of small RNA species in human cells. Northern blotting to detect small RNAs (22 nt in length) using antisense (left panel) and sense (right panel) probes derived from nt 21–40 of RMRP. f, g, Analysis of the termini of the small RNA species identified in (e). Total RNA was incubated with the indicated enzyme (f), or oxidation-β-elimination reactions (g) were performed. Northern blotting was performed with antisense probe. CIP = calf intestinal phosphatase. PNK = polynucleotide kinase. ATP-indicates samples lacking ATP.