FIG 4.

Properties of short RNAs synthesized by TERT RdRP. (A) IP-RdRP assay using a lysate of silkworm pupae overexpressing rTERT, an anti-human TERT MAb (clone 10E9-2), and [α-³²P]UTP. Synthetic RNAs (RNA 1 and RNA 2) were used as templates. The IP-RdRP assay without template RNA was performed as the NC (control). (B) Scheme for the preparation of a small RNA library for deep sequencing analysis of IP-RdRP products. RppH was used to convert 5′ ends of triphosphate to monophosphate. The library was successfully constructed from 5′-triphosphorylated RNAs with RppH treatment. (C) Length distributions of type D sequences in rTERT-RNA 1 and rTERT-RNA 2. IP-RdRP products of rTERT were analyzed by deep sequencing. (D) Length distributions of type D sequences in DMSO-RNA 1 and nocodazole-RNA 1. Short RNA products in the IP-RdRP assay using HeLa cells were analyzed by deep sequencing.