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: ARv7 promotes the escape of PCa cells from AIS. A Left panel, stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with normal medium or CSS medium for 7 days and then subjected SA-β-gal activity assays. Right panel, the expression levels of ARv7 in two cell lines were analyzed using IB. B Stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with DMSO control or 10 μM enzalutamide for 72 h and then subjected to an SA-β-gal activity assay. C Stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with normal medium or CSS medium for 7 days and then subjected to EdU assay. D Stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with DMSO or 10 μM enzalutamide for 72 h and then subjected to EdU assay. E Left panel, stable virus-infected 22Rv1 cells (shctrl or shARv7 or shARv7 transfected with GFP-ARv7) were treated with DMSO control or 40 μM enzalutamide for 72 h and then subjected to SA-β-gal activity assays. Right panel, the expression levels of endogenous and exogenous ARv7 in three groups were analyzed using IB. F Stable virus-infected 22Rv1 cells (shctrl or shARv7 or shARv7 transfected with GFP-ARv7) were treated with DMSO or 40 μM enzalutamide for 72 h and then subjected to EdU assays. G Stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with DMSO or 10 μM enzalutamide for 72 h and then subjected to IB. H Stable virus-infected 22Rv1 cells (shctrl or shARv7) were transfected with empty vector or GFP-ARv7 plasmid for 24 h, treated with 40 μM enzalutamide for another 48 h, and then subjected to IB. I Upper panel, stable virus-infected LNCaP cells (vector or ARv7 OE) were treated with 10 μg/ml CHX for different times as indicated and then harvested for IB. Lower panel, stable virus-infected 22Rv1 cells (shctrl or shARv7) were treated with 10 μg/ml CHX for different times as indicated and then harvested for IB. J HeLa cells were transfected with the indicated plasmids for 48 h, treated with 10 μM MG132 for an additional 6 h, and then harvested for IB and IP using the antibodies indicated in the figure

Article Snippet: Chromatin samples were immunoprecipitated with antibodies against ARv7 (CST, 19672) or a negative control rabbit IgG antibody (CST).

Techniques: Virus, Infection, Plasmid Preparation, Activity Assay, Expressing, Control, EdU Assay, Transfection

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: Outgrowth from AIS is associated with increased levels of ARv7 and decreased levels of p27. A LNCaP cells were cultured in CSS medium for different times as indicated and harvested for the SA-β-gal activity assay. B LNCaP cells were cultured in CSS medium for different times and then harvested for IB. C LNCaP and LNCaP-AI cells were treated with normal medium or CSS medium for 7 days and then subjected to an SA-β-gal activity assay. D LNCaP and LNCaP-AI cells were treated with normal medium or CSS medium for 7 days and then subjected to EdU assay. E LNCaP and LNCaP-AI cells were treated with DMSO or 10 μM enzalutamide for 72 h and then subjected to an SA-β-gal activity assay. F Randomly growing LNCaP and LNCaP-AI cells were harvested for IB analysis against p27

Article Snippet: Chromatin samples were immunoprecipitated with antibodies against ARv7 (CST, 19672) or a negative control rabbit IgG antibody (CST).

Techniques: Cell Culture, Activity Assay, EdU Assay

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: SKP2 is crucial for regulating p27 and AIS. A GSEA analysis of RNA-seq data showing the negative enrichment of the cell-cycle gene signature in ARv7-depleted 22Rv1 cells. B Heat map analysis between two groups of 22Rv1 cells (shctrl or shARv7) shows where expression is high (red) or low (green) for each hallmark cell-cycle gene. C LNCaP cells were treated with 10 μM enzalutamide for 72 h or CSS medium for 7 days and then harvested for IB. D LNCaP cells were treated with 10 μM enzalutamide for 72 h or CSS medium for 7 days and then harvested for qRT-PCR analysis. Data represent the average of three independent experiments ± SD. PSA levels were also checked for measuring the efficacy of ADT-mimicking treatment. E LNCaP cells were cultured in CSS medium for different times and then harvested for IB using an antibody against SKP2. F LNCaP and LNCaP-AI cells were harvested for IB and qRT-PCR analysis. G LNCaP cells were pretreated w/o 1 μM SKP2 inhibitor C1 for 24 h and then cultured with 10 μg/ml CHX for different times as indicated before being harvested for IB. H LNCaP cells were treated with 1 μM SKP2 inhibitor C1 for 24 h and then harvested for the SA-β-gal activity assay. I LNCaP cells were treated with 1 μM SKP2 inhibitor C1 for 24 h and then harvested for EdU assay. J LNCaP cells were treated with 1 μM SKP2 inhibitor C1 for 24 h and then harvested for FACS analysis

Article Snippet: Chromatin samples were immunoprecipitated with antibodies against ARv7 (CST, 19672) or a negative control rabbit IgG antibody (CST).

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Cell Culture, Activity Assay, EdU Assay

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: 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: Chromatin samples were immunoprecipitated with antibodies against ARv7 (CST, 19672) or a negative control rabbit IgG antibody (CST).

Techniques: ChIP-qPCR, Control, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Virus, Infection, Quantitative RT-PCR, Positive Control, Expressing, EdU Assay