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. 2017 Jul 22;7(12):3053–3067. doi: 10.7150/thno.19542

Figure 3.

Figure 3

TGFβ1 represses lncRNA-LET expression in UBC cells. (A) Western blotting showing the levels of p-SMAD2 and total SMAD2 in xenografts treated with or without GEM. (B-C) mRNA levels of lncRNA-LET were measured by qRT-PCR in T24 and 5637 cells treated with or without TGFβ1 or TGFβR1 inhibitor, SB-431542, as well as transfected with control (siNC) or 2 different RNAi of SMAD4 (siSMAD4#1, siSMAD4#2), followed by vehicle or TGFβ1 treatment. (D) Conserved SMAD4 binding element (SBE) in the promoter of human and Rhesus Monkey lncRNA-LET. (E) Relative luciferase activity in T24 cells transfected with plasmids of wild type (pGL3-LET-WT) and SBE deleted (pGL3-LET-ΔSBE) lncRNA-LET promoter, respectively, followed by treatment with vehicle or TGFβ1. (F-G) Sphere numbers (F) and expression of CSC markers (G) in T24 and 5637 cells treated with TGFβ1 alone or TGFβ1 and forced expression of lncRNA-LET. Data are shown as mean ± SD and represent at least two independent experiments with similar results. * P < 0.05, ** P < 0.01, *** P < 0.001 (Student's unpaired two-tailed t-test).