Research Paper Volume 12, Issue 14 pp 14754—14774

Figure 7. DSCAM-AS1 protected DCTPP1 mRNA from degradation mediated by miRNAs. (A) In the mRNA stability assay, Actinomycin D was added to BC cells to prevent cell transcription. mRNA level was depicted at different time points, as measured by PCR. (B) Bioinformatics analysis using a RNA-RNA interaction software (http://rna.informatik.uni-freiburg.de/IntaRNA/Input.jsp) showed tight junctions between DCTPP1 mRNA and DSCAM-AS1 at three areas. (C) There are more than 70 bases from 822 to 935 locus in DCTPP1 mRNA bound to the bases in DSCAM-AS1. The region from 822 to 935 locus locates in the 3’UTR of DCTPP1 mRNA. (D) Many miRNAs, such as miR-3173-5p, miR-874-3p and miR-874-3p, were predicted to bind to DCTPP1 mRNA in the same region. (E) Interaction between DCTPP1 mRNA and DSCAM-AS1 was confirmed by RNA-pull down test. (F) RIP analysis showed that DSCAM-AS1 knockdown increased the enrichment of DCTPP1 mRNA, miR-3173-5p, miR-874-3p and miR-874-3p in the AGO2 protein complex when cell transcription was inhibited by Actinomycin D. (G) As indicated by FISH assay, DSCAM-AS1 knockdown decreased the levels of DCTPP1 mRNA and miR-3173-5p in MCF-7 and T47D cells. *P<0.05 and **P<0.01.