Journal: Endocrinology

Article Title: Induction of Amnion Epithelial Apoptosis by Cortisol via tPA/Plasmin System.

doi: 10.1210/en.2016-1464

Figure Lengend Snippet: Figure 4. Involvement of tPA/plasmin system in the induction of apoptosis by cortisol in human amnion epithelial cells. (A-D) Blockade of cortisol (1 M)-induced increases in cleaved-caspase-3 protein abundance and activity by plasmin inhibitor tranexamic acid (TA, 20 M) (A, n 4), siRNA (50 nM)-mediated knockdown of plasminogen (B, n 5 for protein or n 3 for activity), tPA antibody (1:100) (C, n 5 for protein or n 3 for activity) and siRNA (50 nM)-mediated knockdown of tPA (D, n 5 for protein or n 3 for activity). (E) tPA (1, 5 and 10ng/mL) increased the amounts of cleaved-caspase-3 protein and activity. n 5 for protein or n 4 for activity. The top panels of A-E are the representative immunoblots. (F) tPA (5ng/mL) increased the number of apoptotic cells in human amnion epithelial cells as revealed with TUNEL staining. Scale bar, 10 m. The left panel is the representative image. n 3. Data are the mean SEM. Statistical analysis was performed with one-way ANOVA test (A-E) or paired Student’s t test (F). *P .05, **P .01, ***P .001 vs. control (0) or negative scrambled siRNA control (NC); ##P .01, ###P .001 vs. cortisol or NC cortisol.

Article Snippet: To study the role of plasmin and tPA in the induction of apoptosis by cortisol, the epithelial cells were treated with cortisol (1 M) in the presence or absence of plasmin inhibitor tranexamic acid (20 M, Santa Cruz) or tPA antibody (1:100, Santa Cruz) or small interfering RNA (siRNA) (siRNA, GenePharma, Shanghai, China)-mediated knockdown of plasminogen and tPA.

Techniques: Quantitative Proteomics, Activity Assay, Knockdown, Western Blot, TUNEL Assay, Staining, Control