Journal: JCI Insight

Article Title: E4 engages uPAR and enolase-1 and activates urokinase to exert antifibrotic effects

doi: 10.1172/jci.insight.144935

Figure Lengend Snippet: PBS ( n = 4), bleomycin ( n = 5) (1.2 mU/g), or bleomycin and E4 ( n = 5) (20 μg/mouse) was administered to C57BL/6J male mice. BALF and lung tissues were collected after 14 days. ( A ) Protein levels of uPA and PAI-1 were detected by immunoblotting (upper) and quantitative analysis (lower). ( B ) Activity levels of uPA and PAI-1 in BALF were measured by activity assays. ( C ) Protein levels of uPA and PAI-1 in mouse lung tissue homogenates were detected by immunoblotting. Western blots (upper) and quantitative analysis (lower) are shown. Samples were electrophoresed on the same gel in noncontiguous lanes. Loss of uPAR abrogates the antifibrotic effects of E4. ( D ) Normal lung fibroblasts were transfected with control or uPAR siRNA and treated with Scr or E4 peptide (10 μg/mL) for 72 hours. FN, COL1α1, uPAR, and EGR-1 levels were detected in whole cell lysates (upper), and MMP-1, MMP-3, and PAI-1 were detected in culture media supernatants (lower). GAPDH and Ponceau S stain were used as loading controls for lysates and supernatants, respectively. C, control siRNA. ( E – G ) Plaur –/– mice were treated with PBS ( n = 7), BLM ( n = 10) (1.5 mU/g), or BLM with E4 ( n = 10) (20 μg/mouse). Lung tissues were collected after 14 days. BLM, bleomycin. ( E ) Collagen content quantified by hydroxyproline assay. ( F ) mRNA levels of Fn and Col1a1 were measured relative to the housekeeping gene Gapdh . ( G ) FN, Col1α1, uPA, and PAI-1 protein levels in lung homogenates of Plaur –/– mice detected by immunoblotting. Western blots (left) and graphical presentation of data (right) are shown. β-Actin was used as a loading control. Samples were run in parallel. Statistical analysis was performed using 2-tailed unpaired Student’s t test and 1-way ANOVA as appropriate; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Error bars are mean ± SD.

Article Snippet: Conflict of interest: CFB is an inventor on patents (8507441, 8716232, 9365616, 9556252, 10172923, 10709769, 10844392) for the antifibrotic effects of E4 and has received research funding from iBio Inc., which has licensed the intellectual property from the University of Pittsburgh.

Techniques: Western Blot, Activity Assay, Transfection, Control, Staining, Hydroxyproline Assay

Journal: Cell reports

Article Title: Tumor-initiating stem cell shapes its microenvironment into an immunosuppressive barrier and pro-tumorigenic niche

doi: 10.1016/j.celrep.2021.109674

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: E4 pipette , Rainin , 17014490.

Techniques: Blocking Assay, Purification, Control, Western Blot, Produced, Affinity Purification, Recombinant, Membrane, Transferring, Cell Isolation, DNA Synthesis, Red Blood Cell Lysis, Knock-Out, Stripping, Gene Expression, Software