Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A-C) Immunofluorescence staining and quantification of YAP or TAZ co-stained with PDGFRα in lung sections from human patients diagnosed with pulmonary fibrosis compared with normal adult human lung (n = 3). (D-F) Immunofluorescence staining and quantification of YAP or TAZ co-stained with PDGFRα in wildtype mice lung sections at 7 days post-bleomycin injury compared to saline-treated controls (n = 5). (G-H) Immunoblot analysis of YAP, TAZ, and CTGF on primary mouse lung fibroblasts treated with/without TGFβ1 (10 ng/mL) for 72 hours (n = 4). Quantification of indicated proteins relative to β-actin. The data are represented as the mean ± SEM; comparison by two-tailed unpaired t-test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Immunofluorescence, Staining, Saline, Western Blot, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: Representative immunofluorescence images from lung tissue sections after bleomycin-induced injury stained for DDR2 (yellow), Vimentin (green), YAP (red), TAZ (pink), and Dapi (blue). (A) Human patients diagnosed with pulmonary fibrosis showed increased nuclear presence of YAP or TAZ in lung fibroblasts compared with normal adult human lung. Red arrows indicate the cells that are positive for DDR2 and Vimentin together with YAP or TAZ. (B) Lung fibroblasts showed increased nuclear presence of YAP or TAZ at 7 days after bleomycin treatment. Red arrows indicate the cells that are positive for DDR2 and Vimentin together with YAP or TAZ in bleomycin-induced fibrotic lung.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Immunofluorescence, Staining

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A) Graphical presentation of experimental design illustrating the bleomycin induced lung fibrosis; Both Yap flox/flox ; Taz flox/flox (control) and Col1a2 Cre(ER)T ; Yap flox/flox ; Taz flox/flox ( dKO ) mice were challenged to one dose of intratracheal administration of bleomycin (2.5 mg/kg body weight) or saline with following intraperitoneal injection of tamoxifen (1 mg per kg body weight/day) as indicated time points. Lung tissue and bronchoalveolar lavage fluid (BALF) were harvested at 14 days post-bleomycin injury for further analysis. (B) Enzyme-linked immunosorbent assay (ELISA) to measure the protein release of pro-inflammatory cytokines IL1β, IL6 and TNF-α in BALF collected from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. (C) Real-time qPCR for pro-inflammatory cytokines and chemokines such as Il6 , Il1β , Tnfα, Cx3cl1, and Cx3cr1 ; with Yap/Taz target gene Ctgf using lung tissue RNA isolated from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. (D-E) Immunostaining and quantification of F4/80 positive macrophage infiltration on the lung from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. The data are represented as the mean ± SEM; comparison by two-tailed unpaired t-test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Control, Saline, Injection, Enzyme-linked Immunosorbent Assay, Isolation, Immunostaining, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A-B) Representative images from lung tissue sections after bleomycin-induced injury stained for PDGFRα (green), YAP (red), TAZ (pink), and Dapi (blue). Lung fibroblasts showed increased nuclear presence of YAP and TAZ at 7 days after bleomycin treatment in control mice. In dKO lung, the expression of YAP and TAZ were inactivated in PDGFRα fibroblasts. White arrows indicate the cells that are positive for PDGFRα together with YAP or TAZ in controls lung section; however, in dKO lung white arrows indicate the cells that are positive for PDGFRα only, but negative for YAP or TAZ expression.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Staining, Control, Expressing

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A-B) Sirius Red/Fast Green staining and quantification (presented as Ashcroft score) on lung sections from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. (C) Quantification of hydroxyproline content using lung tissue from control and dKO mice (n = 5-6) treated with saline or bleomycin for 14 days. (D) Real-time qPCR for profibrotic genes Acta2, SM22α, Col1a1, and Periostin on lung tissue from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. (E-F) Immunoblot analysis and quantification of αSMA and collagen I protein using lung tissue lysate from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. Quantification of indicated proteins relative to β-actin. (G-H) Immunofluorescence staining and quantification of αSMA and collagen I in lung sections from control and dKO mice (n = 5) treated with saline or bleomycin for 14 days. Fluorescence intensity was quantified with imageJ software. The data are represented as the mean ± SEM; comparison by two-tailed unpaired t-test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Staining, Control, Saline, Western Blot, Immunofluorescence, Fluorescence, Software, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A) Graphical presentation of experimental design illustrating the bleomycin induced lung fibrosis; Both R26 YAP5SA (control) and Col1a2 Cre(ER)T ; R26 YAP5SA (Yap over expressing mice) were challenged to one dose of intratracheal administration of bleomycin (2.5 mg/kg body weight) or saline with following intraperitoneal injection of tamoxifen (1 mg per kg body weight/day) as indicated time points. Lungs were harvested at 14 days post-bleomycin injury for further analysis. (B) Real-time qPCR for pro-inflammatory cytokines and chemokines such as Il6 , Il1β , Tnfα, Cx3cl1, and Cx3cr1 ; with Yap/Taz target gene Ctgf using lung tissue RNA isolated from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5) treated with saline or bleomycin for 14 days. (C-D) Immunostaining and quantification of F4/80 positive macrophage infiltration on the lung from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5) treated with saline or bleomycin for 14 days. (E-F) Sirius Red/Fast Green staining and quantification (presented as Ashcroft score) on lung sections from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5) treated with saline or bleomycin for 14 days. (G) Quantification of hydroxyproline content using lung tissue from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5-6) treated with saline or bleomycin for 14 days. (H) Real-time qPCR for profibrotic genes Acta2, SM22α, Col1a1, and Periostin on lung tissue from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5) treated with saline or bleomycin for 14 days. (I-J) Immunoblot analysis and quantification of collagen I and αSMA protein levels using lung tissue lysate from R26 YAP5SA and Col1a2 Cre(ER)T ; R26 YAP5SA mice (n = 5) treated with saline or bleomycin for 14 days. Quantification of indicated proteins relative to β-actin. The data are represented as the mean ± SEM; comparison by two-tailed unpaired t-test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Control, Expressing, Saline, Injection, Isolation, Immunostaining, Staining, Western Blot, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: YAP/TAZ activation in fibroblasts coordinates fibrotic remodeling, fibroinflammation, and epithelial dysfunction in pulmonary fibrosis

doi: 10.1101/2025.06.23.661212

Figure Lengend Snippet: (A) Graphical presentation of experimental design; wild-type mice were challenged to one dose of intratracheal administration of bleomycin (2.5 mg/kg body weight) at day 0 followed by intraperitoneal injection of verteporfin (50 mg/kg body weight) or DMSO from day 1 until day 19, at every alternative day as indicated. DMSO was used as vehicle control. Lungs were harvested at 21 days post-bleomycin injury to assess the lung fibrosis. (B-C) Sirius Red/Fast Green staining and quantification (presented as Ashcroft score) on lung sections from wild-type mice (n = 5-6) treated with DMSO or bleomycin with/without verteporfin for 21 days. (D) Quantification of hydroxyproline content using lung tissue from wild-type mice (n = 5-6) treated with DMSO or bleomycin with/without verteporfin for 21 days. (E) Real-time qPCR for profibrotic genes Acta2 , SM22α , Col1a1, P4ha1, Plod2, Periostin and Ccl2 on lung tissue from wild-type mice (n = 5) treated with DMSO or bleomycin with/without verteporfin for 21 days. (F-G) Immunoblot analysis and quantification of Yap, Taz, collagen I and αSMA protein levels using lung tissue lysate from wild-type mice (n = 5) treated with DMSO or bleomycin with/without verteporfin for 21 days. Quantification of indicated proteins relative to β-actin. (H-I) Immunofluorescence staining and quantification of αSMA and collagen I in lung sections from wild-type mice (n = 5) treated with DMSO or bleomycin with/without verteporfin for 21 days. (J-K) Immunostaining and quantification of Pro-SPC+AT2-cells and Hopx+AT1-cells in wild-type mouse lungs (n=5) treated with DMSO or bleomycin with/without verteporfin for 21 days. Fluorescence intensity was quantified with imageJ software. The data are represented as the mean ± SEM; comparison by one-way ANOVA, followed by Tukey’s post-hoc test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.

Article Snippet: Adult human control lung tissues were purchased from Novus Biologicals (Catalogue no. NBP2-77573, NBP2-30182 and NBP2-42696) and human lung tissues diagnosed with fibrotic diseases were purchased from Origene (Catalogue no. CS504154, CS600794 and CS505314).

Techniques: Injection, Control, Staining, Western Blot, Immunofluorescence, Immunostaining, Fluorescence, Software, Comparison