Fig 1: tPA protein labeling with activity-based probe in cell lysates and in malignant ascites samples.A, JHOC5 control cultures and cultures subjected to PLAT knockdown were labeled with the activity-based probe, and then conditioned media were analyzed by Western blotting. Anti-tPA detection (left panel) identified a clear band at the expected size for single-chain tPA (red arrow) in the control sample; no band is detected after PLAT knockdown. Streptavidin detection of the biotin tag (right panel) reveals faint probe-labeling of single-chain tPA in the control sample (red arrow) while this band is absent in the knockdown sample. B, malignant ascites fluid, retrieved from three mice that carried intraperitoneal JHOC5 tumors, was incubated with the activity-based probe and then analyzed by Western blotting. Anti-tPA (left panel) detected a band for single-chain tPA protein; this band was also very strongly detected by streptavidin (right panel), confirming robust probe-labeling of this species in all three samples. tPA, tissue plasminogen activator.
Fig 2: Tumor cell–expressed tissue-type plasminogen activator (tPA) drives malignant phenotype in OCCC.A and B, expression of PLAT, the gene encoding tPA, was efficiently silenced using two independent lentiviral shRNA constructs in OCCC cell lines JHOC5 (A) and JHOC9 (B). C and D, silencing of PLAT significantly impaired tumor cell invasion in Matrigel transwell assays of JHOC5 cells (C) and JHOC9 cells (D) compared to controls. E and F, silencing of PLAT significantly impaired tumor cell proliferation as reflected in viability assays of JHOC5 cells (E) and JHOC9 cells (F) compared to controls. Significance assessed with one-way ANOVA; **p < 0.01, ***p < 0.001, ****p < 0.0001. OCCC, ovarian clear cell carcinoma; tPA, tissue plasminogen activator.
Supplier Page from Abcam for Anti-TPA Tissue Plasminogen Activator antibody