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