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a. Schematic showing AR-p300/CBP DALTAC-1 rewires the domain-domain interactions of the native AR/p300/CBP complex. AR, androgen receptor; NTD, N-terminal domain; CTD, C-terminal domain; BRD, bromodomain; HAT, histone acetyltransferase. b. Schematic illustrating the structure of AR-p300/CBP DALTAC-1 (JZY3032). c. Left: Immunoblot analysis demonstrating stabilization of p300, CBP, and AR, but not <t>BRD4,</t> by DALTAC-1 in a cellular thermal shift assay in VCaP cells. Right: Graphs of relative expression of the indicated proteins in vehicle (dimethyl sulfoxide, DMSO) or DALTAC-1 treated VCaP cells. d. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. e. Representative images of colonies of DMSO, 10 nM DALTAC-1, or 10 nM p300/CBPi+ARi treated 22Rv1 cells. n=3 biologically replicated wells. f. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, Neg-1 (Negative control-1), or Neg-2 (Negative control-2). n=3 biologically replicated wells. g. Dose-response curves and IC50 of Kelly cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. h. Rank-order plot of IC50 values for DALTAC-1 and CBPD-409 across different human normal and cancer cell lines following 5 days of treatment. Models of AR-positive prostate cancer, AR-negative prostate cancer, and non-neoplastic prostatic cells, neuroblastoma cell lines, Ewing sarcoma cell lines, and breast cancer cell lines are highlighted in specific colors. i. Scatterplot illustrating IC50 values of individual cell lines treated with DALTAC-1 (x-axis) and CBPD-409 (y-axis). AR-positive prostate cancer cell lines are highlighted in red. j. Growth curves of LNCaP cells treated with 100 nM DALTAC-1, combined p300/CBPi + ARi, or CBPD-409 under washout or continuous treatment conditions, monitored using Incucyte Live-Cell Analysis. Data represent mean ± standard deviation (SD), n=6 biologically replicated wells. Statistical significance was assessed using two-way ANOVA.
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a. Schematic showing AR-p300/CBP DALTAC-1 rewires the domain-domain interactions of the native AR/p300/CBP complex. AR, androgen receptor; NTD, N-terminal domain; CTD, C-terminal domain; BRD, bromodomain; HAT, histone acetyltransferase. b. Schematic illustrating the structure of AR-p300/CBP DALTAC-1 (JZY3032). c. Left: Immunoblot analysis demonstrating stabilization of p300, CBP, and AR, but not BRD4, by DALTAC-1 in a cellular thermal shift assay in VCaP cells. Right: Graphs of relative expression of the indicated proteins in vehicle (dimethyl sulfoxide, DMSO) or DALTAC-1 treated VCaP cells. d. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. e. Representative images of colonies of DMSO, 10 nM DALTAC-1, or 10 nM p300/CBPi+ARi treated 22Rv1 cells. n=3 biologically replicated wells. f. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, Neg-1 (Negative control-1), or Neg-2 (Negative control-2). n=3 biologically replicated wells. g. Dose-response curves and IC50 of Kelly cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. h. Rank-order plot of IC50 values for DALTAC-1 and CBPD-409 across different human normal and cancer cell lines following 5 days of treatment. Models of AR-positive prostate cancer, AR-negative prostate cancer, and non-neoplastic prostatic cells, neuroblastoma cell lines, Ewing sarcoma cell lines, and breast cancer cell lines are highlighted in specific colors. i. Scatterplot illustrating IC50 values of individual cell lines treated with DALTAC-1 (x-axis) and CBPD-409 (y-axis). AR-positive prostate cancer cell lines are highlighted in red. j. Growth curves of LNCaP cells treated with 100 nM DALTAC-1, combined p300/CBPi + ARi, or CBPD-409 under washout or continuous treatment conditions, monitored using Incucyte Live-Cell Analysis. Data represent mean ± standard deviation (SD), n=6 biologically replicated wells. Statistical significance was assessed using two-way ANOVA.

Journal: bioRxiv

Article Title: Rewiring Oncogenic Transcriptional Complexes with Domain-ALTeration Chimeras (DALTACs) in Prostate Cancer

doi: 10.64898/2026.04.24.720638

Figure Lengend Snippet: a. Schematic showing AR-p300/CBP DALTAC-1 rewires the domain-domain interactions of the native AR/p300/CBP complex. AR, androgen receptor; NTD, N-terminal domain; CTD, C-terminal domain; BRD, bromodomain; HAT, histone acetyltransferase. b. Schematic illustrating the structure of AR-p300/CBP DALTAC-1 (JZY3032). c. Left: Immunoblot analysis demonstrating stabilization of p300, CBP, and AR, but not BRD4, by DALTAC-1 in a cellular thermal shift assay in VCaP cells. Right: Graphs of relative expression of the indicated proteins in vehicle (dimethyl sulfoxide, DMSO) or DALTAC-1 treated VCaP cells. d. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. e. Representative images of colonies of DMSO, 10 nM DALTAC-1, or 10 nM p300/CBPi+ARi treated 22Rv1 cells. n=3 biologically replicated wells. f. Dose-response curves and IC50 of AR-positive VCaP and LNCaP cells treated with DALTAC-1, Neg-1 (Negative control-1), or Neg-2 (Negative control-2). n=3 biologically replicated wells. g. Dose-response curves and IC50 of Kelly cells treated with DALTAC-1, CBPD-409, or p300/CBPi+ARi. n=3 biologically replicated wells. h. Rank-order plot of IC50 values for DALTAC-1 and CBPD-409 across different human normal and cancer cell lines following 5 days of treatment. Models of AR-positive prostate cancer, AR-negative prostate cancer, and non-neoplastic prostatic cells, neuroblastoma cell lines, Ewing sarcoma cell lines, and breast cancer cell lines are highlighted in specific colors. i. Scatterplot illustrating IC50 values of individual cell lines treated with DALTAC-1 (x-axis) and CBPD-409 (y-axis). AR-positive prostate cancer cell lines are highlighted in red. j. Growth curves of LNCaP cells treated with 100 nM DALTAC-1, combined p300/CBPi + ARi, or CBPD-409 under washout or continuous treatment conditions, monitored using Incucyte Live-Cell Analysis. Data represent mean ± standard deviation (SD), n=6 biologically replicated wells. Statistical significance was assessed using two-way ANOVA.

Article Snippet: For immunoblotting, the following antibodies were used: p300 (Invitrogen: MA1-16608); AR (Abcam: ab133273); H2BK20ac (Cell Signaling Technology: 34156S); H2BK16ac (Abcam: ab177427); H3K27ac (Cell Signaling Technology: 8173S); H2B (Active Motif: 39210); H3 (Cell Signaling Technology: 3638S); c-Myc (Cell Signaling Technology: 9402S); c-Myc/N-Myc (Cell Signaling Technology: 13987S); KLK3/PSA (Dako: A0562); NKX3-1 (Cell Signaling Technology:83700S); CCND1 (Abcam: ab16663); Vinculin (Cell Signaling Technology: 18799S); GAPDH (Santa Cruz Biotechnology: sc-47724); BRD4 (Bethyl Laboratories: A700-004CF), HA-Tag (Cell Signaling Technology: 3724S).

Techniques: Western Blot, Thermal Shift Assay, Expressing, Negative Control, Cell Analysis, Standard Deviation