Journal: Nature communications

Article Title: Circadian clock molecule REV-ERBα regulates lung fibrotic progression through collagen stabilization.

doi: 10.1038/s41467-023-36896-0

Figure Lengend Snippet: Fig. 1 | Decreased REV-ERBα protein abundance and increased protein levels of COL1A1 and LOX in IPF lungs compared to healthy control. Healthy control and IPF formalin fixed-paraffin embedded (FFPE) lung samples were purchased from Origene Inc. Healthy controls contained 100% normal lung architecture with 85% alveoli surface area. IPF patient samples contained at least 50% lesion surface area. The protein abundance of REV-ERBα, COL1A1, and LOX were visualized and

Article Snippet: Compared to the control groups, the protein abundanceof REV-ERBαwas decreased in the healthy area (Control: 21.294%vs. healthy area from IPF: 11.296%) Fig. 1 | Decreased REV-ERBα protein abundance and increasedprotein levels of COL1A1 and LOX in IPF lungs compared to healthy control.Healthy control and IPF formalin fixed-paraffin embedded (FFPE) lung samples were purchased from Origene Inc.

Techniques: Quantitative Proteomics, Control

Journal: Nature communications

Article Title: Circadian clock molecule REV-ERBα regulates lung fibrotic progression through collagen stabilization.

doi: 10.1038/s41467-023-36896-0

Figure Lengend Snippet: Fig. 7 | IAV infection induced dysregulation of profibrotic progression exa- cerbated in Rev-erbα Het mice. WT and Rev-erbα Het mice (equal number of male and female) infected (103 PFU/mouse) with IAV for 15 days, and lungs were sepa- rated for RNA/protein isolation, or fixed with 10% formalin for FFPE sections. a The protein abundance of COL1A2, VIM and activated LOX were measured by western blot. Representative blot images were shown. Different targets were run on the same membrane: COL1A2, VIM and activated LOX were probed in the same membrane and β-ACTIN was used as an endogenous control (n = 5–6 mice per group). b The localizations of COL1A1 and LOX were determined by

Article Snippet: Compared to the control groups, the protein abundanceof REV-ERBαwas decreased in the healthy area (Control: 21.294%vs. healthy area from IPF: 11.296%) Fig. 1 | Decreased REV-ERBα protein abundance and increasedprotein levels of COL1A1 and LOX in IPF lungs compared to healthy control.Healthy control and IPF formalin fixed-paraffin embedded (FFPE) lung samples were purchased from Origene Inc.

Techniques: Infection, Isolation, Quantitative Proteomics, Western Blot, Membrane, Control

Journal: Respiratory research

Article Title: Monoclonal enolase-1 blocking antibody ameliorates pulmonary inflammation and fibrosis.

doi: 10.1186/s12931-023-02583-3

Figure Lengend Snippet: Fig. 1 ENO1 is upregulated in fibrotic lungs from human and bleomycin-treated mice. (A, B) IHC staining of ENO1 was performed in commercially avail able human normal (n = 3) and fibrosis (n = 3) lung FFPE sections. (C, D) After intratracheal injection of bleomycin (BLM, 3 mg/kg) (n = 7) or PBS vehicle control (Sham) (n = 4), the mouse lungs were harvested on day 21 for preparation of FFPE sections and subjected to IHC staining of ENO1. Representative pictures (A, C) and quantitative results of ENO1-stained positive areas were shown (B, D). After intratracheal injection of bleomycin (BLM, 3 mg/kg) (n = 4) or PBS vehicle control (Sham) (n = 4), the mouse lungs were harvested on day 21 for preparation of lysates and subjected to Western blotting for ENO1 (E) and the relative densitometry was shown below the representative blot after GAPDH normalization. Cropped blots were shown, and supplementary Fig. S1 presented the full-length blots. Quantitative results were shown by fold change after bleomycin treatment (F). Scale bar, 100 µM. *P < 0.05, **P < 0.01. (A, B) One experiment was performed and each picture or data point was from one human subject. (C-F) Data were representative for two independent experiments. Each picture, data point, or protein band was from one mouse except (F) was shown as mean ± SD

Article Snippet: Three human fibrotic lung FFPE sections (#CS701530, #CS702702, #CS703355) were obtained from OriGene (Rockville, MD, USA).

Techniques: Immunohistochemistry, Injection, Control, Staining, Western Blot

Journal: Respiratory research

Article Title: Monoclonal enolase-1 blocking antibody ameliorates pulmonary inflammation and fibrosis.

doi: 10.1186/s12931-023-02583-3

Figure Lengend Snippet: Fig. 2 Blocking ENO1 ameliorates lung fibrosis in bleomycin-treated mice. After intratracheal injection of 3 mg/kg bleomycin (BLM) (day 0), mice were treated with ENO1 Ab HL217 (10 mg/kg) intravenously on a 6-day interval from day 1. Mouse lungs were harvested on day 14 or 21 for preparation of FFPE sections and subjected to H&E staining (A) and Masson’s trichrome staining (B). Representative pictures on day 21 (A, B) and quantitative results of Ashcroft score on day 21 (C) and inflammation score on day 14 (D) were shown. Body weight change (E), ratio of lung weight versus body weight (F), col lagen content of the lungs (G), and levels of TGF-β in BALF (H) were shown. Scale bar, 100 µM. *P < 0.05, **P < 0.01, ***P < 0.001. Data were representative for two independent experiments. Each picture or data point was from one mouse except (E) was shown as mean ± SEM

Article Snippet: Three human fibrotic lung FFPE sections (#CS701530, #CS702702, #CS703355) were obtained from OriGene (Rockville, MD, USA).

Techniques: Blocking Assay, Injection, Staining