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du145  (ATCC)


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    Structured Review

    ATCC du145
    ITRI-148 induces degradation of both AR and AR-V7 proteins in CRPC cells. (A) Chemical structures of ITRI-148 and its inactive analog. (B) Expression of Flag-tagged AR-V7 in 293T cells following ITRI-148 treatment. Cells transiently expressing Flag-AR-V7 were pretreated with 50 μg/mL cycloheximide for 1 hour, then exposed to ITRI-148 at the indicated concentrations for 8 hours. AR-V7 expression was detected using an anti-Flag antibody. (C) Representative immunoblots showing AR protein levels in VCaP and CWR22Rv1 cells after 48 hours of ITRI-148 treatment. An N-terminal AR antibody was used to detect both full-length AR and LBD-truncated variants (AR-V(ΔLBD)) in CWR22Rv1 cells, while AR-V7 was detected using an AR-V7-specific antibody. DC₅₀ values were calculated from normalized band intensities, and Dₘₐₓ represents the maximal degradation achieved at 10 μM ITRI-148. (D) Immunofluorescence staining of AR after 24 or 48 hours of ITRI-148 treatment, detected using an N-terminal AR antibody. (E) Western blot analysis of AR and AR-V7 expression in cells treated with DMSO (mock), 10 μM ITRI-148, or the inactive analog. (F) Western blot analysis of ectopically expressed wild-type (WT) and mutant Myc-tagged AR in <t>DU145</t> cells following 24 hours of treatment with 10 μM ITRI-148. Quantified AR levels are as indicated.
    Du145, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 8533 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sigma+plot+program+14+5/pmc12661400-49-2-10?v=ATCC
    Average 99 stars, based on 8533 article reviews
    du145 - by Bioz Stars, 2026-08
    99/100 stars

    Images

    1) Product Images from "Oral bioavailable ITRI-148 degrades androgen receptor variants and overcomes antiandrogen resistance in advanced prostate cancer"

    Article Title: Oral bioavailable ITRI-148 degrades androgen receptor variants and overcomes antiandrogen resistance in advanced prostate cancer

    Journal: Neoplasia (New York, N.Y.)

    doi: 10.1016/j.neo.2025.101253

    ITRI-148 induces degradation of both AR and AR-V7 proteins in CRPC cells. (A) Chemical structures of ITRI-148 and its inactive analog. (B) Expression of Flag-tagged AR-V7 in 293T cells following ITRI-148 treatment. Cells transiently expressing Flag-AR-V7 were pretreated with 50 μg/mL cycloheximide for 1 hour, then exposed to ITRI-148 at the indicated concentrations for 8 hours. AR-V7 expression was detected using an anti-Flag antibody. (C) Representative immunoblots showing AR protein levels in VCaP and CWR22Rv1 cells after 48 hours of ITRI-148 treatment. An N-terminal AR antibody was used to detect both full-length AR and LBD-truncated variants (AR-V(ΔLBD)) in CWR22Rv1 cells, while AR-V7 was detected using an AR-V7-specific antibody. DC₅₀ values were calculated from normalized band intensities, and Dₘₐₓ represents the maximal degradation achieved at 10 μM ITRI-148. (D) Immunofluorescence staining of AR after 24 or 48 hours of ITRI-148 treatment, detected using an N-terminal AR antibody. (E) Western blot analysis of AR and AR-V7 expression in cells treated with DMSO (mock), 10 μM ITRI-148, or the inactive analog. (F) Western blot analysis of ectopically expressed wild-type (WT) and mutant Myc-tagged AR in DU145 cells following 24 hours of treatment with 10 μM ITRI-148. Quantified AR levels are as indicated.
    Figure Legend Snippet: ITRI-148 induces degradation of both AR and AR-V7 proteins in CRPC cells. (A) Chemical structures of ITRI-148 and its inactive analog. (B) Expression of Flag-tagged AR-V7 in 293T cells following ITRI-148 treatment. Cells transiently expressing Flag-AR-V7 were pretreated with 50 μg/mL cycloheximide for 1 hour, then exposed to ITRI-148 at the indicated concentrations for 8 hours. AR-V7 expression was detected using an anti-Flag antibody. (C) Representative immunoblots showing AR protein levels in VCaP and CWR22Rv1 cells after 48 hours of ITRI-148 treatment. An N-terminal AR antibody was used to detect both full-length AR and LBD-truncated variants (AR-V(ΔLBD)) in CWR22Rv1 cells, while AR-V7 was detected using an AR-V7-specific antibody. DC₅₀ values were calculated from normalized band intensities, and Dₘₐₓ represents the maximal degradation achieved at 10 μM ITRI-148. (D) Immunofluorescence staining of AR after 24 or 48 hours of ITRI-148 treatment, detected using an N-terminal AR antibody. (E) Western blot analysis of AR and AR-V7 expression in cells treated with DMSO (mock), 10 μM ITRI-148, or the inactive analog. (F) Western blot analysis of ectopically expressed wild-type (WT) and mutant Myc-tagged AR in DU145 cells following 24 hours of treatment with 10 μM ITRI-148. Quantified AR levels are as indicated.

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining, Mutagenesis

    ITRI-148 induces AR degradation through a ubiquitin-proteasome-dependent mechanism. (A) Proximity ligation assay (PLA) detecting the physical interaction between AR and CRBN upon ITRI-148 treatment. 293T cells transiently expressing Flag-AR or Flag-AR∆LBD were treated with DMSO or ITRI-148 for 8 hours, followed by PLA detection using anti-Flag and anti-CRBN antibodies. (B) Ubiquitination of AR and AR∆LBD proteins. 293T cells transfected with Flag-AR or Flag-AR∆LBD and HA-Ub were incubated with ITRI-148 and MG132. AR proteins were immunoprecipitated using anti-Flag, and ubiquitination of the AR proteins was detected by western blotting with anti-HA. (C) Western blot of Flag-AR and Flag-AR∆LBD in DU145 cells treated with ITRI-148 alone or in combination with MG132 (5 μM), MLN4924 (0.5 mM), or lenalidomide (80 μM). Inhibitors were added for 8 or 10 hours after the 24-hour ITRI-148 treatment. AR expression was detected using anti-Flag antibody.
    Figure Legend Snippet: ITRI-148 induces AR degradation through a ubiquitin-proteasome-dependent mechanism. (A) Proximity ligation assay (PLA) detecting the physical interaction between AR and CRBN upon ITRI-148 treatment. 293T cells transiently expressing Flag-AR or Flag-AR∆LBD were treated with DMSO or ITRI-148 for 8 hours, followed by PLA detection using anti-Flag and anti-CRBN antibodies. (B) Ubiquitination of AR and AR∆LBD proteins. 293T cells transfected with Flag-AR or Flag-AR∆LBD and HA-Ub were incubated with ITRI-148 and MG132. AR proteins were immunoprecipitated using anti-Flag, and ubiquitination of the AR proteins was detected by western blotting with anti-HA. (C) Western blot of Flag-AR and Flag-AR∆LBD in DU145 cells treated with ITRI-148 alone or in combination with MG132 (5 μM), MLN4924 (0.5 mM), or lenalidomide (80 μM). Inhibitors were added for 8 or 10 hours after the 24-hour ITRI-148 treatment. AR expression was detected using anti-Flag antibody.

    Techniques Used: Ubiquitin Proteomics, Proximity Ligation Assay, Expressing, Transfection, Incubation, Immunoprecipitation, Western Blot



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