Fig. 7: CCDC719-13 promotes SLC7A11 degradation and modulates ferroptosis in prostate cancer cells.
A, B Western blot analysis of SLC7A11 and GPX4 protein levels in PC3 cells treated with cycloheximide (CHX, 500 nM) to block protein synthesis. Cells were transfected with either control vector or CCDC719-13 overexpression. C Co-immunoprecipitation (Co-IP) assay showing the ubiquitination of SLC7A11 in HEK293 cells co-transfected with Flag-CCDC719-13 and HA-Ub. Cells were treated with MG132 (10 µM) to inhibit proteasomal degradation. D Quantification of SLC7A11 protein levels over time in CHX-treated PC3 cells. Western blot analysis of SLC7A11 protein levels in PC3 (E) and DU145 (F) cells transfected with control or Flag-CCDC719-13 vector, with or without MG132 treatment. Relative cell viability of PC3 (G) and DU145 (H) cells transfected as indicated. Colony formation assay of PC3 (I) and DU145 (J) cells transfected as indicated. Quantification of cell death in PC3 (K) and DU145 (L) cells as indicated. Cell death was measured by flow cytometry and presented as a percentage. M, N Relative ferroptosis levels in PC3 and DU145 cells transfected as indicated were determined by MDA levels. O Representative bioluminescence images of orthotopic BALB/c mice model injected with PC3 cells transfected as indicated. P Quantification of bioluminescence signal from (O), representing tumor burden in the mice. Q Kaplan–Meier survival curve of mice injected with PC3 cells transfected as indicated.
