TGF-β-Induced α-SMA Expression is Mediated by C/EBPβ Acetylation in Human Alveolar Epithelial Cells In Vitro
This study sought to determine whether binding of acetylated C/EBPβ to α-SMA promoter could affect its activity and was essential for EMT and extracellular matrix deposition in IPF using in vitro model. The expression of EMT and C/EBPβ in A549 cells with TGF-β as pulmonary fibrotic model were detected by western blotting and qPCR. Collagen-I expression using ELISA was performed. The luciferase activity was used to examine the activity of C/EBPβ. Knockdown of C/EBPβ was performed by siRNA. We also investigated the effect of deacetylation of C/EBPβ on EMT using SIRT1. The binding ability of C/EBPβ with α-SMA promoter was affirmed via ChIP and EMSA. The relationship of α-SMA and acetylated C/EBPβ was investigated by Co-IP. SiRNA-mediated knockdown of C/EBPβ in A549 cells attenuated TGF-β1-induced myofibroblast differentiation and ECM deposition. The extent of association between acetylated C/EBPβ and α-SMA promoter was dynamically monitored. It was confirmed that deacetylation of C/EBPβ in A549 cells successfully ameliorated TGF-β1-induced EMT, as shown by reduction in α-SMA expression and excessive collagen-I accumulation. The EMT and fibrotic effect of TGF-β1 dependent on acetylated C/EBPβ-mediated regulation of α-SMA gene activity. C/EBPβ acetylation may play a central role in pulmonary fibrosis.
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Posted 17 Sep, 2020
On 06 Oct, 2020
Received 03 Oct, 2020
Received 28 Sep, 2020
On 18 Sep, 2020
On 17 Sep, 2020
Invitations sent on 17 Sep, 2020
On 17 Sep, 2020
On 16 Sep, 2020
On 15 Sep, 2020
On 11 Sep, 2020
TGF-β-Induced α-SMA Expression is Mediated by C/EBPβ Acetylation in Human Alveolar Epithelial Cells In Vitro
Posted 17 Sep, 2020
On 06 Oct, 2020
Received 03 Oct, 2020
Received 28 Sep, 2020
On 18 Sep, 2020
On 17 Sep, 2020
Invitations sent on 17 Sep, 2020
On 17 Sep, 2020
On 16 Sep, 2020
On 15 Sep, 2020
On 11 Sep, 2020
This study sought to determine whether binding of acetylated C/EBPβ to α-SMA promoter could affect its activity and was essential for EMT and extracellular matrix deposition in IPF using in vitro model. The expression of EMT and C/EBPβ in A549 cells with TGF-β as pulmonary fibrotic model were detected by western blotting and qPCR. Collagen-I expression using ELISA was performed. The luciferase activity was used to examine the activity of C/EBPβ. Knockdown of C/EBPβ was performed by siRNA. We also investigated the effect of deacetylation of C/EBPβ on EMT using SIRT1. The binding ability of C/EBPβ with α-SMA promoter was affirmed via ChIP and EMSA. The relationship of α-SMA and acetylated C/EBPβ was investigated by Co-IP. SiRNA-mediated knockdown of C/EBPβ in A549 cells attenuated TGF-β1-induced myofibroblast differentiation and ECM deposition. The extent of association between acetylated C/EBPβ and α-SMA promoter was dynamically monitored. It was confirmed that deacetylation of C/EBPβ in A549 cells successfully ameliorated TGF-β1-induced EMT, as shown by reduction in α-SMA expression and excessive collagen-I accumulation. The EMT and fibrotic effect of TGF-β1 dependent on acetylated C/EBPβ-mediated regulation of α-SMA gene activity. C/EBPβ acetylation may play a central role in pulmonary fibrosis.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6