Fig. 1

TGF-β activates lnc-UTGF transcription via the canonical TGF-β/SMAD signaling pathway. a, b TGF-β increased lnc-UTGF level in a time- and dose-dependent manner. SK-HEP-1 or SNU-449 cells were untreated (-) or treated with 2 ng/ml TGF-β for the indicated time (a) or with the indicated dose of TGF-β for 24 h (b). c Treatment with transcription inhibitor abolished the TGF-β-induced lnc-UTGF expression. Cells were incubated without or with TGF-β or actinomycin-D (ActD) for 6 h. d-f Inhibition of TGFβR1 or simultaneous knockdown of SMAD2/3/4 attenuated the TGF-β-induced lnc-UTGF expression. For d, cells were incubated without or with TGF-β or TGFβR1 inhibitor (SB525334) for 12 h. For e–f, cells transfected with the indicated RNA duplexes were incubated without or with TGF-β for 24 h. iMAX, cells treated with Lipofectamine RNAiMAX without RNA duplexes. NC, cells transfected with negative control RNA duplex. siTGFβR1-1 and siTGFβR1-2, cells transfected with siRNA targeting different sequences of TGFβR1. siSMADs-1 and siSMADs-2, cells transfected with the mixture of siRNAs targeting SMAD2, SMAD3, and SMAD4. Lnc-UTGF level was detected by qPCR analysis, and U6 was used as an internal control. + or −, cells with (+) or without (−) the indicated treatment. Error bars: SEM from at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant