goat polyclonal anti sry sex determining region y box 2 sox2  (R&D Systems)

Journal: STAR Protocols

Article Title: Protocol to study adult neurogenesis in fresh-frozen human hippocampal tissue using an immunofluorescence quantitative approach

doi: 10.1016/j.xpro.2025.104344

Figure Lengend Snippet: Overview of expected outcomes (A) Schematic representation of distinct cell populations related to adult hippocampal neurogenesis (AHN). (B–H) Expected staining with anti- (B) vimentin, (C) sex determining region Y-box 2 (Sox2), (D) phospho-histone 3 (PH3), (E) HuC-HuD, (F) Doublecortin (DCX), (G) polysialylated-neural cell adhesion molecule (PSA-NCAM), and (H) calbindin (CB) antibodies. (I) Schematic representation of the components of the human hippocampal neurogenic niche. (J–M) Expected staining with anti- (J) S100 calcium-binding protein β (S100β), (K) ionized calcium-binding adaptor molecule 1 (Iba1), (L) phosphorylated γH2A.X (γH2A.X) antibodies, and (M) Ulex Europaeus Agglutinin-I (UEA-1). Yellow scale bar: 20 μm. White scale bar: 10 μm. Yellow triangles indicate positive cells. White triangles highlight the morphology of cells labelled for each marker.

Article Snippet: Goat polyclonal anti-SRY (sex-determining region Y)-box 2 (Sox2) (1:500) , R and D Systems , Cat# AF2018; RRID: AB_355110.

Techniques: Staining, Binding Assay, Marker

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Western blot panels of both non tumorigenic and malignant prostate cells for AR, FOXA1, and SOX2. Blots for β-actin were used as an internal loading control. B) Western blot panels of AR-negative PCa cell lines for AR, FOXA1, SOX2. Blots for β-actin were used as an internal loading control. C-D) Western blots show successful SOX2 shRNA knockdown at 48h in CWR-R1 and 22Rv1 adenocarcinoma cells. Protein bands were quantified using Empiria Studio. E-F) Normalized survival plots of CWR-R1 and 22Rv1 cells after doxycycline induction of shRNA targeting SOX2 or a non-specific scrambled shRNA (sh-SCR). Data points represent the mean normalized survival ± SD; each point is normalized to its own untreated control (without DOX) at each time interval. G-H) Bar graphs represent normalized survival of both CWR-R1 and 22Rv1 cells 7 days after dox-induction of shRNA. I-J) Western blot show successful SOX2 knockdown at 48h via shRNA in LASCPC-01 and siRNA in NCI-H660 NEPC cells. Protein bands were quantified using Empiria Studio. K) Normalized survival plots of LASCPC-01 NEPC cells after doxycycline induction of shRNA targeting SOX2 or a non-specific scrambled shRNA (sh-SCR). Data points represent the mean normalized survival ± SD; each point is normalized to its own untreated control (without DOX) at each time interval. L) Survival plot of NCI-H660 NEPC cells treated with siRNAs targeting SOX2. Data points represent mean luminescence +/- SD. M) Bar graph represents normalized survival of LASCPC-01 NEPC cells 7 days after dox-induction of shRNA. N) Bar graph represents raw survival of NCI-H660 NEPC cells 12 days after siRNA transfection. Data points represent mean luminescence +/- SD.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Western Blot, Control, shRNA, Knockdown, Transfection

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A-B) Western blots show SOX2 protein levels are unaffected by induction of scrambled shRNA at 48h in CWR-R1 and 22Rv1 adenocarcinoma cells. Protein bands were quantified using Empiria Studio. C-E) Growth curves of CWR-R1, 22Rv1, and LASCPC-01 PCa cells across 7 days after dox-induction of shRNAs targeting either SOX2 or scrambled control. Data points represent mean luminescence +/- SD.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Western Blot, shRNA, Control

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) All candidate TFs within 20bp of SOX2 peak in CWR-R1 cells. The e-value is the lowest p-value of any spacing of the secondary motif times the number of secondary motifs; it estimates the expected number of random secondary motifs that would have the observed minimum p-value or less. FOXA1 identified in red. The FOXA1-SOX2 motif enriched by ChIP-seq shows a potential direct interaction on the chromatin. B-F) Gene network signature analyses in PCa patient tumors using the Algorithm for Linking Activity Networks (ALAN) model. Datasets from publicly available sources are depicted across contexts of prostate cancer (GTEX, TCGA, SU2C, and Adeno and NEPC). Gene network signature for FOXA1 (orange) is visualized compared to the gene networks of AR (blue) and SOX2 (red) in each dataset where the median ALAN profile score for each gene network is noted with a black line. Overall network analysis is represented as positive for a median ALAN profile score above 0 and negative for a median ALAN profile score below 0. G) Proximity ligation assay (PLA) measures co-localization (red) of SOX2 and FOXA1 CWR-R1 castration-resistant prostate adenocarcinoma cells. H) Reciprocal co-immunoprecipitation of SOX2 and FOXA1 in CWR-R1 cells. I) Co-immunoprecipitation of FOXA1 with SOX2 in the NEPC cell line NCI-H660. J) Split nano-luciferase complementation reporter assay (Nano-BiT) demonstrating a specific interaction between SOX2 and FOXA1 in HEK293T cells. Tagged AR and FOXA1 were shown to interact as a positive control, and co-transfection of tagged AR and SOX2 were a negative control. Data points represent mean luminescence +/- SD. K) Lentiviral SOX2 overexpression (OE) ablates the AR-FOXA1 Nano-BiT interaction in HEK293T cells. Data points represent mean luminescence +/- SD. L) Lentiviral AR overexpression (OE) ablates the SOX2- FOXA1 Nano-BiT interaction in HEK293T cells, specifically in ARSI conditions. Data points represent mean luminescence +/- SD. M) Nano-BiT in prostate adenocarcinoma LNCaP cells demonstrates strong SOX2-FOXA1 interaction. No significant changes were observed across different AR-signaling contexts. Data points represent mean luminescence +/- SD. N) Western blots of CWR-R1 protein lysate from cells that underwent AR-pathway modulation. Protein bands were quantified using Empiria Studio.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: ChIP-sequencing, Activity Assay, Proximity Ligation Assay, Immunoprecipitation, Luciferase, Reporter Assay, Positive Control, Cotransfection, Negative Control, Over Expression, Western Blot

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Western blots show successful overexpression of AR and SOX2 protein after lentiviral transduction of HEK293T cells. LNCaP and LASCPC-01 cells are included as controls. B) Brightfield (top row) and GFP (bottom row) microscopy images of the lentivirus-transduced, puromycin-selected HEK293T cells. C) Split nano-luciferase complementation reporter assay (Nano-BiT) optimization shows a specific interaction between SOX2 and FOXA1 in HEK293T cells, regardless of the terminal tag location. Tagged AR and FOXA1 were shown to interact as a positive control, and single-BiT controls were included as a negative control. Data points represent mean luminescence +/- SD. D) Nano-BiT in AR-overexpressing HEK293T cells shows no change in SOX2 interaction with clinically relevant FOXA1 mutants. Data points represent mean luminescence +/- SD.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Western Blot, Over Expression, Transduction, Microscopy, Luciferase, Reporter Assay, Positive Control, Negative Control

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Binding motif enrichment near AR, FOXA1, and SOX2 peaks in CWR-R1 cells. Both MEME and STREME motif discovery analyses were included. Full lists of motifs discovered can be found in Supplementary Table 1. B) C) Venn diagram of AR, FOXA1, and SOX2 ChIP-seq peaks in CWR- R1 cells grown in whole media. Shared regions represent called peaks with >= 1bp of overlap. D) Venn diagram of AR, FOXA1, and SOX2 potential target genes. CWR-R1 ChIP-seq peaks were analyzed using Cistrome-GO to identify genes that had AR, FOXA1, and/or SOX2 transcription factor peaks less than 10kb from a TSS and an adjusted regulatory potential (RP) score of >0.01. E-H) Gene Ontology (GO) pathway analyses for the unique and shared AR, FOXA1, and SOX2 candidate target genes in CWR-R1. GO enrichment was performed using Enrichr. I-K) Track plots of AR, FOXA1, and SOX2 ChIP-seq peaks in CWR-R1 undergoing AR pathway modulation. Peaks visualized using IGV and mapped to the human hg38 chromosome build.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Binding Assay, ChIP-sequencing

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near AR binding sites alone in CWR- R1 cells. GO enrichment was performed using Enrichr; data shown as -log( p ). B) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near FOXA1 binding sites alone in CWR-R1 cells. GO enrichment was performed using Enrichr; data shown as -log( p ). C) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near AR/FOXA1 co-bound sites without SOX2 in CWR-R1 cells. GO enrichment was performed using Enrichr; data shown as -log( p ). D) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near AR/SOX2/FOXA1 co-bound sites in CWR- R1 cells. GO enrichment was performed using Enrichr; data shown as -log( p ).

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Binding motif enrichment near FOXA1 and SOX2 peaks in NCI-H660 NEPC cells. Both MEME and STREME motif discovery analyses were included. Full lists of motifs discovered can be found in Supplementary Table 1. B) Venn diagram of SOX2 and FOXA1 ChIP-seq peaks in NCI-H660 cells grown in complete 5% FBS HITES media. Shared regions represent called peaks with >= 1bp of overlap. C) Venn diagram of SOX2 and FOXA1 potential target genes. NCI-H660 ChIP-seq peaks were analyzed using Cistrome-GO to identify genes that had AR, FOXA1, and/or SOX2 transcription factor peaks less than 10kb from a TSS and an adjusted regulatory potential (RP) score of >0.01. D-F) Gene Ontology (GO) pathway analyses for the unique and shared FOXA1 and SOX2 candidate target genes in NCI- H660. GO enrichment was performed using Enrichr. G-N) Track plots of SOX2 and FOXA1 ChIP- seq peaks near oncogenic/NEPC genes in NCI-H660. Peaks visualized using IGV and mapped to the human hg38 chromosome build.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Binding Assay, ChIP-sequencing

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near FOXA1 binding sites alone in NCI-H660 NEPC cells. GO enrichment was performed using Enrichr; data shown as -log( p ). B) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near SOX2 binding sites alone in NCI-H660 NEPC cells. GO enrichment was performed using Enrichr; data shown as -log( p ). C) Full list of significantly enriched ( p <= 0.05) GO Biological Processes for candidate genes of near SOX2/FOXA1 co-bound sites a in NCI-H660 NEPC cells. GO enrichment was performed using Enrichr; data shown as -log( p ).

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A-F) Track plots of SOX2 and FOXA1 ChIP-seq peaks near oncogenic/NEPC genes in NCI-H660. Peaks visualized using IGV and mapped to the human hg38 chromosome build. G) Relative mRNA expression of SOX2 and ROR1 across PCa cell lines normalized to β-actin. Data are represented as the fold change (2 -ΔΔCT ) ± SEM. H-J) Track plots of AR, FOXA1, and SOX2 co-bound genes in CWR-R1 and NCI-H660. Peaks visualized using IGV and mapped to the human hg38 chromosome build.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: ChIP-sequencing, Expressing

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A-D ) Track plots of AR, FOXA1, and SOX2 co-bound genes in CWR-R1 and NCI-H660. Peaks visualized using IGV and mapped to the human hg38 chromosome build. E) Heatmap of differential gene expression in Control vs. SOX2 KO CWR-R1 cells. Data represent transcripts per million (TPM) values of RNA-seq triplicates. F) UpSet plot of AR and SOX2 ChIP-seq peaks in CWR-R1 cells. Shared regions represent called peaks with >= 1bp of overlap. G) Venn Diagram of AR and SOX2 ChIP-seq peaks in CWR-R1 cells. Shared regions represent called peaks with >= 1bp of overlap. H) Venn diagram of AR and SOX2 potential target genes. CWR-R1 ChIP-seq peaks were analyzed using Cistrome-GO to identify genes that had AR and/or SOX2 binding peaks less than 10kb from a TSS and an adjusted regulatory potential (RP) score of >0.01. I) Gene Ontology (GO) pathway analyses for the invariant AR and SOX2 candidate target genes in CWR-R1. GO enrichment was performed using Enrichr. J) Relative SOX2 and ROR1 mRNA expression in NCI-H660 cells following SOX2 siRNA knockdown normalized to β-actin. Data are represented as the fold change (2 -ΔΔCT ) ± SEM. K) The chosen sequence of the SOX2/FOXA1 co-bound region (peak summit) of the ROR1 gene used for computational modeling of the SOX2/FOXA1 interaction. L) Track plot of SOX2/FOXA1 co-binding in the ROR1 gene body. M) Computational modeling using the Chai-1 build of AlphaFold-3 to predict the structure of SOX2 and FOXA1 (UniProt IDs: P48431 and P55317, respectively) co-bound to DNA containing a single FOX motif within the ROR1 gene. N) Structure prediction of SOX2 and FOXA1 co-bound to DNA containing both an HMG-box and FOX motif within the ROR1 gene.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Gene Expression, Control, RNA Sequencing, ChIP-sequencing, Binding Assay, Expressing, Knockdown, Sequencing

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A-D ) Heatmaps of transcription factor binding in CWR- R1 and NCI-H660 cells. The vertical axis of each heatmap represents the top 1000 transcription factor binding sites by peak score in both adenocarcinoma and NEPC. E) Nested Venn diagram of the shared SOX2/FOXA1 candidate target genes (from and ) in both CWR-R1 and NCI-H660. We find 130 potentially SOX2/FOXA1 co-regulated genes across cell lines. F-H ) Gene Ontology (GO) pathway analyses for the unique and shared CWR-R1 and NCI-H660 candidate SOX2/FOXA1 co-regulated genes. GO enrichment was performed using Enrichr.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) UMAP visualization of prostate cancer cell populations from 13 castration-resistant patients from the Human Metastatic Prostate (HMP) dataset. B) UMAP visualization of the same 13 patients categorized by histological type: adenocarcinoma and NEPC. C) NEPC signature analysis applied to CRPC patients. D-F) Expression analyses of AR , FOXA1 , and SOX2 across the same UMAP coordinates G) Expression levels of genes ( AR , FOXA1 , SOX2 ) and pathway signatures (AR, NEPC scores) are shown through a bubble plot as organized by sample. H) Western blot panels of malignant prostate cancer cells for AR, FOXA1, SOX2, and the FGFR-family of proteins. Blots for β-actin were used as an internal loading control. I) UpSet plot of SOX2 ChIP-seq peaks in CWR-R1, NCI-H660, PrEC, WA-01 and NCCIT cells. Shared regions represent called peaks with >= 1bp of overlap. J) Venn Diagram SOX2 ChIP-seq peaks in CWR-R1, NCI-H660, PrEC, WA-01 and NCCIT cells. Shared regions represent called peaks with >= 1bp of overlap. K) Venn diagram of SOX2 potential target genes. SOX2 ChIP-seq peaks from all cell lines were analyzed using Cistrome-GO to identify genes that SOX2 binding peaks less than 10kb from a TSS and an adjusted regulatory potential (RP) score of >0.01. We find zero genes that are commonly bound by SOX2 across the different cell contexts. L) Binding motif enrichment near SOX2 peaks in PrEC, CWR-R1, and NCI-H660 cells. Both MEME and STREME motif discovery analyses were included. Full lists of motifs discovered can be found in Supplementary Table 1. M) Binding motif enrichment near SOX2 peaks in pluripotent WA-01 and NCCIT cells. Both MEME and STREME motif discovery analyses were included. Full lists of motifs discovered can be found in Supplementary Table 1 .

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Expressing, Western Blot, Control, ChIP-sequencing, Binding Assay

Journal: bioRxiv

Article Title: SOX2 utilizes FOXA1 as a heteromeric transcriptional partner to drive proliferation in therapy-resistant prostate cancer

doi: 10.1101/2025.07.18.664790

Figure Lengend Snippet: A) Expression analyses of ASCL1 and NEUROD1 across the same CRPC patient UMAP coordinates from the HMP dataset in . B) Expression analyses of SOX2 and ASCL1 across the same CRPC patient UMAP coordinates from the HMP dataset in ) Scatter plot showing the relationship between SOX2 and ASCL1 expression in the Beltran (Adeno) dataset. D) Scatter plot showing the relationship between SOX2 and ASCL1 expression in the Beltran (NEPC) dataset. E) Violin plot showing gene expression changes over time for ASCL1 alongside SOX2 . Violin plot expression levels were obtained from the in vivo temporal model of NEPC trans-differentiation [Pan-small cell neuroendocrine cancer (PARCB)] and are displayed across different time points, highlighting temporal patterns and co-expression relationships between target genes and SOX2 . F) Violin plot from the PARCB model showing gene expression changes over time for NEUROD1 alongside SOX2 . G) Violin plot from the PARCB model showing gene expression changes over time for AR alongside SOX2 . H) Scatter plot showing the relationship between SOX2 expression and Basal Activity Score the Beltran (Adeno) dataset. I) Scatter plot showing the relationship between SOX2 expression and Neuroendocrine Activity Score the Beltran (Adeno) dataset. J) Scatter plot showing the relationship between SOX2 expression and Luminal Activity Score the Beltran (Adeno) dataset. K) Scatter plot showing the relationship between SOX2 expression and AR Activity Score the Beltran (Adeno) dataset. L) Graph ranking Spearman correlation coefficients for select genes with respect to Basal Activity Score profiles in the Beltran (Adeno) dataset. Genes like SOX2 and FGFR1 show significant positive correlations, whereas genes such as AR, FOXA1, and NKX3-1 are negatively correlated with Basal Activity Score.

Article Snippet: SOX2 was then immunoprecipitated with Dynabeads Protein G using beads conjugated to polyclonal goat anti-SOX2 antibody (R&D Systems, Cat: AF2018) or the species-matched goat IgG control antibody (R&D Systems, Cat: AB-108-C).

Techniques: Expressing, Gene Expression, In Vivo, Activity Assay