anti foxa1  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: BPTF regulates androgen receptor activity by enhancing chromatin accessibility and stabilizing the AR-FOXA1 interaction

doi: 10.1038/s41467-025-67329-9

Figure Lengend Snippet: A Co-IP of AR with BPTF in Rv1 cells. B Co-IP of HA-AR with the Flag-tagged BPTF fragments in 293 T cells. C Co-IP of HA-AR with Flag-tagged BPTF C-terminal fragment (F6 + 7), but not with its BRD-deleted version (F6 + 7−BRD), in 293 T cells. D Co-IP of myc-tagged BPTF C-terminal fragment (F7) with Flag-tagged AR fragments (N-terminal transactivation domain N-TAD, DNA-binding domain DBD, Ligand-binding domain LBD, wild type WT) in 293 T cells. E Co-IP of BPTF with AR following R1881 treatment in C4-2 cells. F Co-IP of BPTF with AR following enzalutamide (ENZ) treatment in C4-2 cells. G Heatmap showing BPTF ChIP-seq peaks. CUT&RUN ChIP-seq was performed on Rv1 cells using a BPTF antibody or an IgG control antibody. H Genomic distribution of BPTF peaks. I HOMER motif analysis of BPTF peaks showing enrichment of FOXA1, androgen response element (ARE), and AR half-site motifs. J Heatmap of AR ChIP-seq peaks. CUT&RUN ChIP-seq was performed on Rv1 cells using an AR antibody or an IgG control antibody. K Genomic distribution of AR peaks. L HOMER motif analysis of AR peaks showing enrichment of FOXA1, ARE, and AR half-site motifs. M Heatmap showing the overlap between BPTF peaks and AR peaks. N Genome browser track images showing BPTF and AR peaks at the promoter and enhancer regions of representative AR target genes. H3K27ac HiChIP data from Rv1 cells highlight potential enhancer regions for NKX3-1 and SLC45A3 , which form loops with their corresponding promoter region. O, P ChIP-qPCR analysis of BPTF ( O ) and AR ( P ) at promoter or enhancer regions of KLK2 , KLK3 , NKX3−1, and SLC45A3 in Rv1 cells. Q ChIP-qPCR analysis of BPTF and AR at KLK3 enhancer (left) and NKX3−1 promoter (right) following CRISPRa-mediated upregulation of BPTF in Rv1 cells. CUT&RUN ChIP-seq ( G – N ) were generated from two biological replicates. All other data ( A – F , O – Q ) are representative of three independent biological replicates. Data are presented as mean ± SD ( O – Q ). Statistical significance was determined using a two-tailed unpaired Student’s t-test ( O , P) ; One-way ANOVA (two-sided) with Tukey’s multiple comparison test ( Q ). Source data are provided as a file.

Article Snippet: Antibodies were obtained from the following sources: BPTF (ABE24, ABE1966, MABE443) and AR (06-680) from EMD Millipore (Burlington, MA); c-Myc (#9402) and HA-tag (#3724) from Cell Signaling Technology (Danvers, MA); SMARCA1 (13-2005), FOXA1 (13-2001), H3K27ac (13-0059), H3K4me1 (13-0040), and Rabbit IgG control (13-0042) from EpiCypher (Durham, NC); Myc (sc-40) from Santa Cruz Biotechnology (Dallas, TX); Flag (F7425, F3165) and actin (A5441) from Sigma-Aldrich (St. Louis, MO); Cas9 (61957) and RNA Pol II (39497) from Active Motif (Carlsbad, CA); Mouse IgG control (02-6300) and fluorescent secondary antibodies (A-21109, A-21058, A-11375) from Invitrogen (Waltham, MA); and fluorescent TrueBlot secondary antibodies (18-4416-32, 18-4417-32) from Rockland Immunochemicals (Pottstown, PA).

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Ligand Binding Assay, ChIP-sequencing, Control, HiChIP, ChIP-qPCR, Generated, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: BPTF regulates androgen receptor activity by enhancing chromatin accessibility and stabilizing the AR-FOXA1 interaction

doi: 10.1038/s41467-025-67329-9

Figure Lengend Snippet: A Heatmap of RNA-seq data showing that FOXA1 KD in Rv1 cells decreases the expression of a subset of BPTF-dependent AR target genes. B Co-IP of FOXA1 with BPTF in Rv1 cells. C Co-IP of HA-FOXA1 with Flag-tagged BPTF fragments (F3, F5, and F6) in 293 T cells. D Reduced co-IP of FOXA1 with AR following BPTF KD in Rv1 cells. E Venn diagram showing the shared ChIP-seq peaks of BPTF, AR and FOXA1. F HOMER motif analysis showing enrichment of FOXA1 and ARE motifs within the shared ChIP-seq peaks of BPTF, AR, and FOXA1. Heatmap ( G ) and profile plots ( H ) showing that FOXA1 KD in Rv1 cells reduces signal intensities of FOXA1, BPTF, and AR peaks at BPTF-dependent AR-binding sites. I Track images showing reduced FOXA1, BPTF, and AR peak signals at the representative AR target genes upon FOXA1 KD in Rv1 cells. Heatmap ( J ) and profile plots ( K ) showing that BPTF KD in Rv1 cells has minimal impact on FOXA1 peak intensity at BPTF-dependent AR-binding sites. Heatmap ( L ) and profile plots ( M ) showing that AR KD has little effect on BPTF peak intensity at BPTF-dependent AR-binding sites. RNA-seq data were generated from three biological replicates ( A ). CUT&RUN ChIP-seq data ( E – M ) were generated from two biological replicates. Data are representative of three independent biological replicates ( B – D ). Source data are provided as a file.

Article Snippet: Antibodies were obtained from the following sources: BPTF (ABE24, ABE1966, MABE443) and AR (06-680) from EMD Millipore (Burlington, MA); c-Myc (#9402) and HA-tag (#3724) from Cell Signaling Technology (Danvers, MA); SMARCA1 (13-2005), FOXA1 (13-2001), H3K27ac (13-0059), H3K4me1 (13-0040), and Rabbit IgG control (13-0042) from EpiCypher (Durham, NC); Myc (sc-40) from Santa Cruz Biotechnology (Dallas, TX); Flag (F7425, F3165) and actin (A5441) from Sigma-Aldrich (St. Louis, MO); Cas9 (61957) and RNA Pol II (39497) from Active Motif (Carlsbad, CA); Mouse IgG control (02-6300) and fluorescent secondary antibodies (A-21109, A-21058, A-11375) from Invitrogen (Waltham, MA); and fluorescent TrueBlot secondary antibodies (18-4416-32, 18-4417-32) from Rockland Immunochemicals (Pottstown, PA).

Techniques: RNA Sequencing, Expressing, Co-Immunoprecipitation Assay, ChIP-sequencing, Binding Assay, Generated

Journal: Nature Communications

Article Title: BPTF regulates androgen receptor activity by enhancing chromatin accessibility and stabilizing the AR-FOXA1 interaction

doi: 10.1038/s41467-025-67329-9

Figure Lengend Snippet: A , B Predicted 3D interaction model of BPTF-BRD (cyan) and AR-LBD (green) generated by AlphaFold2. The BPTF-BRD pocket region is highlighted in dark blue, while H3, H4, and H12 of the AR co-factor binding groove are highlighted in magenta, orange, and yellow, respectively. The model is visualized as a ribbon diagram ( A ) or a solid surface ( B ). C Superimposed structure of AU1-docked BPTF-BRD with the predicted BPTF-BRD and AR-LBD interaction model. AU1 is highlighted in red, the BPTF-BRD pocket in dark blue, and H3, H4, and H12 of the AR co-factor binding groove in magenta, orange, and yellow, respectively. The superimposed model is visualized as a ribbon diagram. D Western blot showing BPTF, AR, and FOXA1 protein levels after AU1 treatment in Rv1 cells. E Decreased co-IP of AR with BPTF following AU1 treatment in Rv1 and C4-2 cells. F qRT-PCR analysis showing that AU1 treatment decreases mRNA levels of representative AR target genes in Rv1 and C4-2 cells. G , H IC 50 determination of AU1 in PCa cells using colony formation assays. Colony formation assays evaluating the effects of DMSO, AU1 (IC 50 concentration), enzalutamide (ENZ, 5 μM), and combination treatment (ENZ + AU1) on PCa cells. Example images ( I ) and quantification ( J ) of colony formation are shown. Data are representative of three independent biological replicates ( D – J ). Data are presented as mean ± SD ( F , G ). Statistical significance was determined using one-way ANOVA (two-sided) with Tukey’s multiple comparison test ( F , J ). Source data are provided as a file.

Article Snippet: Antibodies were obtained from the following sources: BPTF (ABE24, ABE1966, MABE443) and AR (06-680) from EMD Millipore (Burlington, MA); c-Myc (#9402) and HA-tag (#3724) from Cell Signaling Technology (Danvers, MA); SMARCA1 (13-2005), FOXA1 (13-2001), H3K27ac (13-0059), H3K4me1 (13-0040), and Rabbit IgG control (13-0042) from EpiCypher (Durham, NC); Myc (sc-40) from Santa Cruz Biotechnology (Dallas, TX); Flag (F7425, F3165) and actin (A5441) from Sigma-Aldrich (St. Louis, MO); Cas9 (61957) and RNA Pol II (39497) from Active Motif (Carlsbad, CA); Mouse IgG control (02-6300) and fluorescent secondary antibodies (A-21109, A-21058, A-11375) from Invitrogen (Waltham, MA); and fluorescent TrueBlot secondary antibodies (18-4416-32, 18-4417-32) from Rockland Immunochemicals (Pottstown, PA).

Techniques: Generated, Binding Assay, Western Blot, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Concentration Assay, Comparison

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Verification of mRNA and protein level expression in FOXA1 siRNA-treated ovarian cancer cells. ( a ) FOXA1 mRNA expression after FOXA1 siRNA transfection. ( b ) Quantification of FOXA1 protein expression. ( c ) Representative Western blot images of FOXA1 and β-actin after FOXA1 siRNA transfection.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Expressing, Transfection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Cell proliferation analysis to silence of FOXA1 in ovarian cancer cells. FOXA1 silencing in SK-OV3 and HEYA8 ovarian cancer cell lines reduces cell proliferation compared to control. (* p < 0.01).

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Cell migration, invasion, and wound healing assay to silence of FOXA1 in ovarian cancer cells. ( a ) FOXA1 si decreased the migration ability of ovarian cancer cells (Left: SK-OV3, Right: HEYA8). ( b ) FOXA1 si decreased the invasion ability of ovarian cancer cells (Left: SK-OV3, Right: HEYA8). ( c ) Representative images of the wound healing assay at 0, 12, 24, 36, and 48 h time points (Left: SK-OV3, Right: HEYA8). ( d ) The quantitative evaluation and statistical analysis of wound area percentage in wound healing assay measured by Image J software. Results are expressed as mean ± SD of five experiments (Left: SK-OV3, Right: HEYA8).

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Migration, Wound Healing Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Comparison of EMT gene expression after treatment with FOXA1 siRNA. ( a ) Changes in EMT gene expression in HEYA8. ( b ) Changes in EMT gene expression in SK-OV3. ( c ) Schematic representation of EMT and MET processes and their key molecular markers.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Comparison, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Reduction in chemoresistance following FOXA1 Gene Knockdown. ( a ) Determining IC50 values using carboplatin. ( b ) Atezolizumab cytotoxicity test in SK-OV3 and HEY A8 cells. ( c , d ) Cell viability of SK-OV3 and HEYA8 ovarian cancer cells following carboplatin treatment with or without atezolizumab. ( e , f ) Cell viability of SK-OV3 and HEYA8 ovarian cancer cells following carboplatin treatment with or without FOXA1 knockdown. ( g , h ) Cell viability of SK-OV3 and HEYA8 ovarian cancer cells following combined treatment with carboplatin and atezolizumab, with or without FOXA1 knockdown. ( i , j ) Comparison of cell viability in FOXA1-silenced SK-OV3 and HEYA8 cells treated with carboplatin alone or in combination with atezolizumab.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Knockdown, Comparison

Journal: International Journal of Molecular Sciences

Article Title: FOXA1 in Ovarian Cancer: A Potential Therapeutic Target to Enhance Immunotherapy Efficacy

doi: 10.3390/ijms27031194

Figure Lengend Snippet: Immunohistochemical analysis of FOXA1 expression in ovarian cancer patient tissue samples. ( a ) High expression of FOXA1. ( b ) Low expression of FOXA1. ( c ) Malignant tumor patients stratified by stage (I–IV) ( p < 0.05). ( d ) Patients under fifty years old stratified into normal, benign, and tumor groups ( p < 0.05). ( e ) Patients under fifty years old stratified by benign vs. tumor only ( p < 0.05).

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies for FOXA1 (Santa Cruz, Dallas, TX, USA, sc-101058, 1:1000) and β-actin (Santa Cruz, Dallas, TX, USA, sc-47778, 1:1000).

Techniques: Immunohistochemical staining, Expressing