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TCF7L2 is a dependency in CRPC-WNT. A, Growth data measured by live-cell imaging (IncuCyte S3) upon transfection of indicated siRNA or plasmid (EV or rescue). Immunoblot of indicated proteins at 72 hours after transfection. GAPDH served as loading control. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. ***, P < 0.001. Data are representative of n = 2 independent experiments. B, Spheroid formation measured by live-cell imaging (IncuCyte SX5) upon transfection of indicated siRNA. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are representative of n = 2 independent experiments. C, IHC and staining intensity of TCF7L2 on indicated organoids upon treatment with 1 µmol/L A858 or 1 µmol/L A947 for 24 hours. Violin plot from TCF7L2 staining intensity analyzed using two-way ANOVA. ****, P < 0.0001. Scale bars, 100 µm (MSK-PCa16) and 50 µm (WCM1078). OD, optical density. D, TOPFlash TCF/LEF reporter assay measured after 48 hours upon treatment with indicated drugs in WCM1078 sublines. FOPFlash served as a negative reporter control. Data are presented as mean values ± SEM after normalization to internal Renilla control and analyzed using a paired Student t test. ***, P < 0.001; ****, P < 0.0001. Data are representative of n = 2 independent experiments.

Journal: Cancer Research

Article Title: A Double-Negative Prostate Cancer Subtype Is Vulnerable to SWI/SNF-Targeting Degrader Molecules

doi: 10.1158/0008-5472.CAN-25-2928

Figure Lengend Snippet: TCF7L2 is a dependency in CRPC-WNT. A, Growth data measured by live-cell imaging (IncuCyte S3) upon transfection of indicated siRNA or plasmid (EV or rescue). Immunoblot of indicated proteins at 72 hours after transfection. GAPDH served as loading control. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. ***, P < 0.001. Data are representative of n = 2 independent experiments. B, Spheroid formation measured by live-cell imaging (IncuCyte SX5) upon transfection of indicated siRNA. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are representative of n = 2 independent experiments. C, IHC and staining intensity of TCF7L2 on indicated organoids upon treatment with 1 µmol/L A858 or 1 µmol/L A947 for 24 hours. Violin plot from TCF7L2 staining intensity analyzed using two-way ANOVA. ****, P < 0.0001. Scale bars, 100 µm (MSK-PCa16) and 50 µm (WCM1078). OD, optical density. D, TOPFlash TCF/LEF reporter assay measured after 48 hours upon treatment with indicated drugs in WCM1078 sublines. FOPFlash served as a negative reporter control. Data are presented as mean values ± SEM after normalization to internal Renilla control and analyzed using a paired Student t test. ***, P < 0.001; ****, P < 0.0001. Data are representative of n = 2 independent experiments.

Article Snippet: After selection with blasticidin, cells were transduced with internal control Renilla luciferase (Rluc) lentivirus (BPS Biosciences, 79565-G).

Techniques: Live Cell Imaging, Transfection, Plasmid Preparation, Western Blot, Control, Staining, Reporter Assay