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. 2019 Mar 5;41:670–682. doi: 10.1016/j.ebiom.2019.02.057

Fig. 2.

Fig. 2

TGF-β1 mediates bleomycin-induced up-regulation of miR-4739 by enhancing the pri-miR-4739 promoter activity. (a) Human PMCs were treated with or without bleomycin (0.2 μg/ml) for 24 h after which the levels of TGF-β1 in the cultured medium were detected by ELISA. n = 3, *P < 0.05 versus control (Student's t-test). (b) Human PMCs were treated with TGF-β1 (5 ng/ml) for 48 h after which miR-4739 RT-qPCR was performed. n = 8, *P < 0.05 versus control (Student's t-test). (c) Human PMCs were treated with bleomycin (0.2 μg/ml) with or without SB431542 (inhibitor of ALK5, type I TGF-β receptor) for 48 h. Then the levels of miR-4739 were detected by RT-qPCR. n = 6, *P < 0.05 versus control; #P < 0.05 versus bleomycin group (ANOVA test). (d) HEK 293 T cells were transfected with empty control vector or miR-4739 promoter plasmid, then the effect of Smad3 on pri-miR-4739 promoter was evaluated. All groups were transfected with Smad4 expression vector and a control renilla vector. n = 4, *P < 0.05 versus empty control vector; #P < 0.05 versus miR-4739 promoter plasmid (ANOVA test). (e-f) Human PMCs were transfected with miR-4739 promoter plasmid or empty control vector. After 24 h, the cells were treated with bleomycin (0.2 μg/ml) or TGF-β1 (0.5 ng/ml), the activity of the pri-miR-4739 promoter was detected. All groups were transfected with a control renilla expression. n = 4, *P < 0.05 versus empty control vector; #P < 0.05 versus miR-4739 promoter plasmid (ANOVA test). (g-h) Human PMCs were treated with 0.2 μg/ml bleomycin (g, n = 4) or 0.5 ng/ml TGF-β1 (h, n = 6) for 24 h, then the pri-miR-4739 level was detected. *P < 0.05 versus control (Student's t-test).