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Fig. 2 | Molecular Medicine

Fig. 2

From: TGF-β-induced α-SMA expression is mediated by C/EBPβ acetylation in human alveolar epithelial cells

Fig. 2

Up-regulation of C/EBPβ is involved in TGF-β-induced EMT. a Real-time PCR analysis of C/EBPβ mRNA levels in A549 cells treated with 0−20 ng/mL TGF-β for 24 h. b C/EBPβ mRNA levels increased in a time-dependent manner of TGF-β (10 ng/mL) treatment (0−48 h) in A549 cells. c TGF-β increased C/EBPβ protein levels in A549 cells in a concentration- (0, 1, 2.5, 5, 10, and 20 ng/mL) dependent manner. d C/EBPβ expression increased gradually by 7.05 ± 0.81 folds by 24 h when TGF-β treatment was prolonged. e, f Quantification of western blots were calculated according to grey analysis. g Luciferase activity assay was performed to assess C/EBPβ promoter activity. Generation of reporter gene constructs controlled by the human α-SMA gene promoter. Reporter constructs were generated by inserting fragments of the α-SMA gene promoter into a luciferase reporter vector pGL4.10. The location of the C/EBPβ-binding motif is as indicated. Relative to the control, the activity of C/EBPβ-luciferase increased with TGF-β concentration and peaked at 10 ng/mL by 2.74 ± 0.42-fold. h TGF-β up-regulated C/EBPβ-luciferase activity. Traces represent three experiments with similar results. Means ± SEM of data are shown. Asterisk (*) denotes significant difference (P < 0.05)

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