Fig. 5
From: Identification of a TGF-β/SMAD/lnc-UTGF positive feedback loop and its role in hepatoma metastasis
Lnc-UTGF interacts with the mRNAs of SMAD2 and SMAD4 and enhances their stability. a–b Silencing lnc-UTGF shortened the half-life of mature mRNAs of SMAD2 and SMAD4. Cells transfected with the indicated RNA duplexes were treated with actinomycin-D (ActD) for the indicated time, followed by qPCR analysis. U6 was used as an internal control. c Complementary base-pairing between lnc-UTGF and SMAD2/4 mRNAs. The putative complementary sequences were predicted by BLAST (https://blast.ncbi.nlm.nih.gov/). d–e Lnc-UTGF was physically associated with the mRNAs of SMAD2 and SMAD4. For d, the SK-UTGF and SK-S1m-UTGF sublines were applied to S1m-tagged RNA affinity purification assays. The indicated RNAs in the S1m-pull-down precipitates and in the input were detected by qPCR. The RNA level of the pull-down product was corrected by that in the input. The mean value of the adjusted RNA level in the pull-down product of the SK-UTGF group was set as relative RNA level 1. For e, the RNA pulled down by biotin-labeled lnc-UTGF or by biotin-labeled antisense RNA of lnc-UTGF (UTGF-AS, negative control) were examined by qPCR. U6, negative control. Error bars: SEM from three independent experiments. *, P < 0.05; **, P < 0.01; ns, not significant
