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. 2016 Dec 10;8(3):5092–5110. doi: 10.18632/oncotarget.13867

Figure 10. Sp1 transcription factor is required for both CUG2 and TGF-β transcription.

Figure 10

A. Production of TGF-β protein was detected with immunoblotting with an anti-TGF-β antibody after running of SDS-PAGE under a reduced condition. Production of TGF-β1 protein in the culture media from A549-CUG2 and BEAS-CUG2 cells was compared to that from their control cell media with a modified sandwich ELISA using Au nanoparticles. The assay was repeated in triplicate. The results shown are the average of triplicate wells and error bars indicate SD. (*; p< 0.05, ***; p< 0.001). Expression of TGF-β1 transcripts from A549-CUG2 and BEAS-CUG2 cells was compared to that from their control cells with qRT-PCR. The assay was repeated triplicate. Each assay was performed in triplicate and error bars indicate SD. (**; p< 0.01, ***; p< 0.001) B-D. A549-Vec, A549-CUG2, BEAS-Vec, or BEAS-CUG2 cells were transfected with TGF-β promoter vectors (phTG1, 5, 6, 7, and 7-4) or CUG2 promoter vectors (F961 and F961-94) in the absence and presence of TGF-β (5 ng/mL). At 48 h post-transfection, luciferase enzyme activities were measured in the transfected cell lysates. Transfection efficiency was normalized with the β-galactosidase reporter vector, pGK-β-gal. The assay was repeated in triplicate. The results shown are the average of triplicate wells. Error bars indicate SD. (*; p< 0.05,**; p<0.01, ***; p< 0.001)