Journal: Science Advances

Article Title: Control of airway basal stem cell–mediated lung repair by TGF-β signaling

doi: 10.1126/sciadv.adz1519

Figure Lengend Snippet: ( A ) Schematic diagram of iBMP7-BCs transplantation into BLM-damaged lungs of immune-deficient NOD-SCID mice. ( B ) Immunostaining detected pSmad2 and pSmad3 in lung tissue at different times following BLM injury. DAPI: nuclear counterstain. ( C ) pSmad2 + and pSmad3 + cells were quantified by ImageJ ( n = 6 technical replicates from three independent mice, one-way ANOVA with Dunnett’s test). ( D ) Time-series direct observations of iBMP7-BCs transplantation using a stereomicroscope. ( E ) Immunostaining for HuLamin and GFP in lung sections after iBMP7-BCs transplantation. HuLamin: lamin A/C, a human-specific nuclear antigen. DAPI: nuclear counterstain. ( F ) The relative GFP intensity was quantified by ImageJ ( n = 3 mice per group). ( G ) Immunofluorescence of BMP7-Flag and GFP expression at 7 days after transplantation (14 dpi) of iBMP7-BCs into BLM-damaged lung. DAPI: nuclear counterstain. ( H ) Axial plane micro-CT images of mouse fibrosis at day 14 following treatment with saline, Ctrl-BCs, or iBMP7-BCs. Lower panels: representative reconstructed 3D images. Red: pulmonary injury area (including intrapulmonary airways); gray: normal lung tissue. Right panel: quantitative analysis of pulmonary injury volume based on CT images ( n = 3 mice per group). ( I ) Collagen deposition was assessed by hydroxyproline content in lung tissue ( n = 4 mice per group). ( J ) qPCR analysis of Col1a1 , Col3a1 , Fn1 , and Acta2 mRNA expression levels in mouse lungs 14 days after treatment with saline, Ctrl-BCs, or iBMP7-BCs ( n = 4 mice per group; two technical replicates were performed for each mouse). ( K ) Arterial blood gas analysis was performed on the arterial blood of mice treated with saline, Ctrl-BCs, or iBMP7-BCs for 14 days ( n = 4 mice per group). All data are represented as the means ± SEM. One-way ANOVA with Tukey’s test was used for (H) to (K). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. ns, not significant.

Article Snippet: The mouse lung was injured by intratracheally instilling with a 3 U/kg body weight of BLM (Selleckchem, Houston, US) 7 days before transplantation.

Techniques: Transplantation Assay, Immunostaining, Immunofluorescence, Expressing, Micro-CT, Saline

Journal: RSC Advances

Article Title: Structure-based design of orthosteric and allosteric CCR2 inhibitors for potential IPF therapy

doi: 10.1039/d5ra05026j

Figure Lengend Snippet: CCR2 Expression and Pathological Validation in IPF Progression. (A–C) In silico analysis based on patient data from dataset GSE70866 . (A) The violin plot depicts CCR2 gene expression levels in the normal control and IPF groups ( GSE70866 ) (* P < 0.1). (B) The survival analysis plot outlines survival probabilities over time for groups with high and low CCR2 expression. (C) The ROC curve demonstrates the diagnostic performance of CCR2 in identifying pulmonary fibrosis. (D–J) In vivo validation using a bleomycin-induced pulmonary fibrosis mouse model. (D) Representative images of H&E and Masson's trichrome staining of lung tissues 4 weeks after intratracheal instillation of BLM in mice. (E) Pathological scoring results of H&E-stained lung tissues at week 4 (** P < 0.01). (F) Analysis of collagen area in lung tissues at week 4 (*** P < 0.001). (G) Hydroxyproline content in lung tissue of bleomycin (BLM)-induced pulmonary fibrosis mice and control mice. The hydroxyproline level was significantly increased in BLM-treated mice compared to controls (*** P < 0.001). (H) RT-qPCR analysis of CCR2 expression levels in BLM-induced idiopathic pulmonary fibrosis (*** P < 0.001). (I and J) Western blot analysis of CCR2 expression levels in lung tissues of each group (** P < 0.01).

Article Snippet: Mice in the experimental group were anesthetized with 1% sodium pentobarbital (50 mg kg −1 ), and a single dose of 5 mg kg −1 bleomycin (BLM) solution (Selleck, USA) was administered intratracheally.

Techniques: Expressing, Biomarker Discovery, In Silico, Gene Expression, Control, Diagnostic Assay, In Vivo, Staining, Quantitative RT-PCR, Western Blot