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Figure 4 | BMC Complementary and Alternative Medicine

Figure 4

From: Research on the efficacy of Celastrus Orbiculatus in suppressing TGF-β1-induced epithelial-mesenchymal transition by inhibiting HSP27 and TNF-α-induced NF-κB/Snail signaling pathway in human gastric adenocarcinoma

Figure 4

The role of HSP27 in COE-mediates inhibition of EMT and NF-κB/Snail signal pathway. (A) Overexpression of HSP27 was performed by transfected with the recombinant retroviral expression plasmid and the cells were selected by flow cytometry for GFP + cells. Nearly 100% transduction efficiency was achieved as shown by fluorescent microscope. (B) Expression of HSP27, E-cadherin, N-cadherin, Vimentin were analyzed by western blot in SGC-7901 cells that were transfected with control (empty) or HSP27 vectors or treated with TGF-β1 (10 ng/mL). β-actin was served as an internal control of protein level. The relative density was normalized to β-actin, which was determined by densitometric analysis. Values are expressed as means±SD of three independent experiments. **P <0.01, compared with control. ##P <0.01, compared to MIGR1 group. (C) The overexpression HSP27 cell line and control cell line were treated with or without COE (20 μg/mL) for 1 h followed by TGF-β1 (10 ng/mL) for 24 h, and the protein expression of HSP27, E-cadherin, N-cadherin, Vimentin were detected by western blot analyses. β-actin was served as an internal control of protein level. (D) Cell invasion and migration were analyzed by transwell assay. The quantitative data was presented as means±SD of three independent experiments. (E) The overexpression HSP27 cell line and control cell line were treated with or without COE (20 μg/mL) for 1 h followed by TNF-α (10 ng/mL) for 24 h, and the protein expression of IκBα, p-IκBα, NF-κB p65 and Snail were detected by western blot analyses. β-actin and H3 were served as an internal control of protein level. **P <0.01 compared with the respective untreated control (MIGR1 or MIGR1-HSP27). ##P <0.01 compared COE-treated MIGR1 group compared with COE-treated MIGR1-HSP27 group.

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