Journal: Translational Oncology
Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization
doi: 10.1016/j.tranon.2025.102626
Figure Lengend Snippet: The functional GTSF1/PIWIL4 complex mediates piR-43452-induced suppression of LRP1 . A. Validation of GTSF1 overexpression in UM-UC-3 and J82 cells. B. Efficiency of GTSF1 knockdown in UM-UC-3 and J82 cells, and the corresponding expression level of PIWIL4.C. RNA immunoprecipitation using an anti-HA antibody in 3HA-GTSF1-overexpressing HEK-293T cells, followed by qPCR, shows enhanced enrichment of both piR-43452 and LRP1 mRNA in the HA immunoprecipitates compared to the IgG control. D. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and GTSF1-overexpressing plasmid.E. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and siGTSF1. F. Interaction of PIWIL4 protein and GTSF1 protein detected with Co-IP. G. Efficiency of PIWIL4 knockdown in UM-UC-3 and J82 cells, and the corresponding expression level of GTSF1.H. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and siPIWIL4. (mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001).
Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).
Techniques: Functional Assay, Biomarker Discovery, Over Expression, Knockdown, Expressing, RNA Immunoprecipitation, Control, Transfection, Plasmid Preparation, Co-Immunoprecipitation Assay