Journal: Redox Biology
Article Title: Hydrogen sulfide attenuates cigarette smoke-induced airway remodeling by upregulating SIRT1 signaling pathway
doi: 10.1016/j.redox.2019.101356
Figure Lengend Snippet: NaHS inhibited EMT in CS-exposed mouse lungs. After CS exposure for 12 weeks, mice were euthanized, and lung tissues were collected. (A) Western blot was used to analyze the protein levels of E-cadherin, CK18, Fibronectin, vimentin, FSP-1, α-SMA, Snail, MMP-2, MMP-9 and MMP-12 in the lung tissues. (B) Densitometric analysis of E-cadherin, CK18, Fibronectin, vimentin, FSP-1, α-SMA, Snail, MMP-2, MMP-9 and MMP-12 in the immunoblots using β-actin as the internal reference. (C) The expression of E-cadherin and vimentin in the lungs were visualized using immunofluorescence double staining. (D, E) Western blot was used to analyze the protein levels of TGF-β1, TGFβR1 and p-Smad3 in mouse lungs. Data are presented as mean ± SEM of 6 mice/group, *P < 0.05, **P < 0.01, significantly different from control group; # P < 0.05, ## P < 0.01, significantly different from CS + Saline group.
Article Snippet: The antibodies used in this study include: anti-matrix metalloproteinase (MMP)-2 (Cat# 10373-2-AP), anti-MMP-9 (Cat# 10375-2-AP), anti-MMP-12 (Cat# 22989-1-AP), anti-fibronectin (Cat# 15613-1-AP), anti-TGF-β1 (Cat# 21898-1-AP) and anti-β-actin (Cat# 66009-1-Ig) polyclonal antibodies were purchased from Proteintech (Chicago, IL, USA); anti-Cytokeratin (CK)18 (Cat# GB11232), anti-collagen 1 (Cat# GB11022-2) and anti-collagen 3 (Cat# GB11023) polyclonal antibodies were purchased from Servicebio (Wuhan, China); anti-fibroblast specific protein (FSP)1 (Cat# A1631), and anti-superoxide dismutase (SOD)2 (Cat# A1340) antibodies were purchased from abclonal (Wuhan, China); anti-E-cadherin (Cat# 14472), anti-vimentin (Cat# 5741), anti-Snail (Cat# 3879), anti-SIRT1 (Cat# 8469), anti-p-Smad3 (Cat# 9520), and anti-Smad3 (Cat# 9523) antibodies were purchased from Cell Signaling Technology (CA, USA); anti-TGF-β1 receptor type Ⅰ (TGFβR1) monoclonal antibody (Cat# AHO1552) was purchased from Thermo Fisher Scientific (Boston, MA, USA); anti-α-SMA antibody (Cat# ab32575) and the HRP-labeled Goat Anti-Rabbit/Mouse IgG (H + L) were purchased from Abcam Biotechnology (Cambridge, MA, USA).
Techniques: Western Blot, Expressing, Immunofluorescence, Double Staining
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Cell Signaling Technology Inc manufactures this product 
![NaHS repressed cigarette smoke extract (CSE)-induced EMT and collagen deposition in human bronchial epithelial 16HBE cells. (A) 16HBE cells were treated with 3% CSE and different concentrations of NaHS (100, 200, or 400 μM) for 48 h. The protein levels of E-cadherin, fibronectin and α-SMA was analyzed by Western blot. (B–D) Densitometric analysis of proteins of interest in the immunoblots using β-actin as the internal reference. (E) Immunofluorescence for E-cadherin was performed on human 16HBE cells treated with and without 3% CSE in the presence of 400 μM NaHS for 48 h. (F–H) Western blot was used to detect collagen 1 and collagen 3 levels. (I–K) Western blot was used to analyze the protein levels of TGF-β1 and p-Smad3. Data are presented as mean ± SEM of at least three independent experiments, **P < 0.01, significantly different from untreated cells [3%CSE (-) and NaHS (-)]; #P < 0.05, ##P < 0.01, significantly different from cells treated with CSE only.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4091/pmc06854091/pmc06854091__gr3.jpg)
