Journal: BMC biology
Article Title: ARv7 promotes the escape of prostate cancer cells from androgen deprivation therapy-induced senescence by mediating the SKP2/p27 axis.
doi: 10.1186/s12915-025-02172-4
Figure Lengend Snippet: Fig. 5 ARv7 mediates AIS in an SKP2-dependent manner. A Integrative Genomics Viewer (IGV) image showing the enrichment for the indicated factor at SKP2 and UBE2C in 22Rv1 cells, ATAC-seq data was applied to show the area of open chromatin. B ChIP-qPCR of ARv7 and IgG control at the promoter site in 22Rv1 cells. Data represent the average of three independent experiments ± SD. C HEK293T cells were co-transfected with SKP2 reporter plasmid, Renilla plasmid, and w/o GFP-ARv7 plasmid for 48 h, then cells were subjected to luciferase assay. Relative luciferase activity was calculated after normalized to Renilla luciferase unit. D Stable virus-infected LNCaP cells (vector or ARv7 OE) were harvested for IB. E Stable virus-infected 22Rv1 cells (shctrl or shARv7) were harvested for IB. F Stable virus-infected LNCaP cells (vector or ARv7 OE) were harvested for qRT-PCR analysis. G Stable virus-infected 22Rv1 cells (shctrl or shARv7) were harvested for qRT-PCR analysis. H Upper panel, PC-3 cells were transfected with empty vector or GFP-ARv7 for 48 h and harvested for IB; LNCaP cell lysate was analyzed together as a positive control for AR-FL expression. Lower panel, PC-3 cells were transfected with empty vector or GFP-ARv7 for 48 h and harvested for qRT-PCR analysis. I Stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with normal medium or CSS medium for 7 days then treated w/o 1 μM SKP2 inhibitor C1 for another 24 h before being harvested for EdU assay
Article Snippet: SKP2 inhibitor C1, cycloheximide, MG132, and enzalutamide were purchased from TargetMol (Boston, MA, USA), while puromycin was purchased from MedChemExpress (Monmouth Junction, NJ, USA).
Techniques: ChIP-qPCR, Control, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Virus, Infection, Quantitative RT-PCR, Positive Control, Expressing, EdU Assay