Journal: International Journal of Molecular Sciences
Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk
doi: 10.3390/ijms27041841
Figure Lengend Snippet: IL-17A induces p53, PAI-1, Cav1 and apoptosis, and CSP or CSP7 inhibits IL-17A-induced apoptosis in AECs. ( A ) AECs isolated from uninjured WT and p53 −/− mice were treated with IL-17A (0–100 ng/mL) for 24 h, and the lysates were immunoblotted for p53, PAI-1, Cl./Cas-3 and β-actin. ( B ) WT mice received saline or IL-17A intranasally (0–3 μg in 50 μL saline), as previously described . AECs were isolated 24 h later, and lysates were immunoblotted for p53 and Cl./Cas-3 and β-actin. ( C ) WT mice were exposed to saline or IL-17A (1 μg), as described in ( B ). Lysates of AECs isolated 24 h after treatment with IL-17A were immunoblotted for ACp53, p53, Sirt1, Cav1 and apoptosis. ( D ) AECs isolated from WT mice were treated with PBS or IL-17A (100 ng/mL). Two hours later, IL-17A-stimulated cells were treated with or without CSP or CP (10 µM) and analyzed for p53, PAI-1 and apoptosis by Western blotting (WB). ( E ) AECs isolated from WT mice exposed to IL-17A (100 ng/mL) were treated with CSP, its overlapping deletion fragments, or control peptide (CP) for 24 h in vitro. The lysates were analyzed for ACp53, p53, PAI-1, Sirt1 and Cl. Cas-3 by WB. ( F ) miR-34a fl/fl mice and miR-34acKO mice lacking miR-34a in AECs were exposed to IL-17A (1 μg) with or without CSP7 or CP. Control mice were exposed to saline. Lysates from AECs extracted from these mice were analyzed for p53, PAI-1, Sirt1, Cav1 and apoptosis by WB. ( G ) Lung homogenates or lysates of AECs extracted from WT mice exposed to IL-17A with or without CSP7 or CP were subjected to WB for PP2Ac, pATM and ATM kinase. ( H ) Lung homogenates of IL-17A-exposed mice treated with or without CSP7 or CP were immunoprecipitated (IP) using anti-Cav1 antibody and immunoblotted for p53, mdm2, and PP2Ac. ( I ) Lung homogenates of WT mice exposed to saline or IL-17A with or without CSP7 were analyzed for MPO activity.
Article Snippet: 17 , PP2Ac , CST , 2259S , 1:1000 , .
Techniques: Isolation, Saline, Western Blot, Control, In Vitro, Immunoprecipitation, Activity Assay
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