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Santa Cruz Biotechnology pc3 nuclear extract

A) His-tag affinity pull-down validation of hits using <t>PC3</t> nuclear extract which contains native EWS. Purified His-ERG was used as bait and ERG protein is shown by Ponceau stain (bottom). EWS signal is shown by EWS immunoblot (top). 62.5 μM of indicated Hit compounds were used. B) His-tag affinity pull-down as in (A) except using 60, 45, and 30 μM of Hit B. Two independent experiments are shown (EXP1, top; EXP2, bottom). C) Cell viability assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. D) Cell migration assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. E) Clonogenic growth assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. F) IC50 determination by migration assay using RWPE1-ERG cells after treatment with DMSO or indicated compound concentrations in μM. The IC50, indicated by the point in which the red dashed line crosses the cell migration curve, is estimated to be ~9 μM. G) IC50 determination by luciferase assay. Luciferase expression is driven by an ETS-motif-containing enhancer element and firefly expression is normalized to renilla expression. IC50, indicated by the point in which the red dashed line crosses the relative luciferase curve, is estimated to be ~9 μM. H) Immunoblot of ERG and EWS from VCaP cells after treatment with 45 μM of indicated compound or DMSO for 24 hr. Tubulin (TUB) is used as a loading control. I) qRTPCR of known ERG target genes after treatment with 45 μM of indicated compound or DMSO for 24 hr in VCaP cells.

Pc3 Nuclear Extract, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pc3+nuclear+extracts/PC-3+nuclear+extract/pmc07485968-48-3-6
Average 93 stars, based on 73 article reviews

pc3 nuclear extract - by Bioz Stars, 2026-10

93/100 stars

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1) Product Images from "A high-throughput screen identifies inhibitors of the interaction between the oncogenic transcription factor ERG and the cofactor EWS"

Article Title: A high-throughput screen identifies inhibitors of the interaction between the oncogenic transcription factor ERG and the cofactor EWS

Journal: PLoS ONE

doi: 10.1371/journal.pone.0238999

A) His-tag affinity pull-down validation of hits using PC3 nuclear extract which contains native EWS. Purified His-ERG was used as bait and ERG protein is shown by Ponceau stain (bottom). EWS signal is shown by EWS immunoblot (top). 62.5 μM of indicated Hit compounds were used. B) His-tag affinity pull-down as in (A) except using 60, 45, and 30 μM of Hit B. Two independent experiments are shown (EXP1, top; EXP2, bottom). C) Cell viability assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. D) Cell migration assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. E) Clonogenic growth assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. F) IC50 determination by migration assay using RWPE1-ERG cells after treatment with DMSO or indicated compound concentrations in μM. The IC50, indicated by the point in which the red dashed line crosses the cell migration curve, is estimated to be ~9 μM. G) IC50 determination by luciferase assay. Luciferase expression is driven by an ETS-motif-containing enhancer element and firefly expression is normalized to renilla expression. IC50, indicated by the point in which the red dashed line crosses the relative luciferase curve, is estimated to be ~9 μM. H) Immunoblot of ERG and EWS from VCaP cells after treatment with 45 μM of indicated compound or DMSO for 24 hr. Tubulin (TUB) is used as a loading control. I) qRTPCR of known ERG target genes after treatment with 45 μM of indicated compound or DMSO for 24 hr in VCaP cells.

Figure Legend Snippet: A) His-tag affinity pull-down validation of hits using PC3 nuclear extract which contains native EWS. Purified His-ERG was used as bait and ERG protein is shown by Ponceau stain (bottom). EWS signal is shown by EWS immunoblot (top). 62.5 μM of indicated Hit compounds were used. B) His-tag affinity pull-down as in (A) except using 60, 45, and 30 μM of Hit B. Two independent experiments are shown (EXP1, top; EXP2, bottom). C) Cell viability assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. D) Cell migration assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. E) Clonogenic growth assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. F) IC50 determination by migration assay using RWPE1-ERG cells after treatment with DMSO or indicated compound concentrations in μM. The IC50, indicated by the point in which the red dashed line crosses the cell migration curve, is estimated to be ~9 μM. G) IC50 determination by luciferase assay. Luciferase expression is driven by an ETS-motif-containing enhancer element and firefly expression is normalized to renilla expression. IC50, indicated by the point in which the red dashed line crosses the relative luciferase curve, is estimated to be ~9 μM. H) Immunoblot of ERG and EWS from VCaP cells after treatment with 45 μM of indicated compound or DMSO for 24 hr. Tubulin (TUB) is used as a loading control. I) qRTPCR of known ERG target genes after treatment with 45 μM of indicated compound or DMSO for 24 hr in VCaP cells.

Techniques Used: Biomarker Discovery, Purification, Staining, Western Blot, Viability Assay, Cell Migration Assay, Growth Assay, Migration, Luciferase, Expressing, Control

Lysis:

Article Title: Phosphorylation of the oncogenic transcription factor ERG in prostate cells dissociates polycomb repressive complex 2, allowing target gene activation
Article Snippet: .. To the bead-bound ETS proteins, ∼14 μg of PC3 nuclear extracts (catalog no. sc-2152; Santa Cruz Biotechnology) were added and rotated for 1–2 h. The beads were washed four times with Nonidet P-40 lysis buffer, and bead-bound proteins were eluted using SDS protein loading dye. ..

Article Title: Interaction with ZMYND11 mediates opposing roles of Ras-responsive transcription factors ETS1 and ETS2
Article Snippet: .. With the bead bound ETS proteins, ∼14 μg of PC3 nuclear extracts (sc-2152, Santa Cruz Biotechnology) along with 100 μg of BSA were rotated for 1–2 h. Beads were washed four times with NP-40 lysis buffer and proteins eluted using sodium dodecyl sulphate (SDS) protein loading dye. ..

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Journal: PLoS ONE

Article Title: A high-throughput screen identifies inhibitors of the interaction between the oncogenic transcription factor ERG and the cofactor EWS

doi: 10.1371/journal.pone.0238999

Figure Lengend Snippet: A) His-tag affinity pull-down validation of hits using PC3 nuclear extract which contains native EWS. Purified His-ERG was used as bait and ERG protein is shown by Ponceau stain (bottom). EWS signal is shown by EWS immunoblot (top). 62.5 μM of indicated Hit compounds were used. B) His-tag affinity pull-down as in (A) except using 60, 45, and 30 μM of Hit B. Two independent experiments are shown (EXP1, top; EXP2, bottom). C) Cell viability assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. D) Cell migration assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. E) Clonogenic growth assay using RWPE1-ERG cells after treatment with 45 μM of indicated compound or DMSO. F) IC50 determination by migration assay using RWPE1-ERG cells after treatment with DMSO or indicated compound concentrations in μM. The IC50, indicated by the point in which the red dashed line crosses the cell migration curve, is estimated to be ~9 μM. G) IC50 determination by luciferase assay. Luciferase expression is driven by an ETS-motif-containing enhancer element and firefly expression is normalized to renilla expression. IC50, indicated by the point in which the red dashed line crosses the relative luciferase curve, is estimated to be ~9 μM. H) Immunoblot of ERG and EWS from VCaP cells after treatment with 45 μM of indicated compound or DMSO for 24 hr. Tubulin (TUB) is used as a loading control. I) qRTPCR of known ERG target genes after treatment with 45 μM of indicated compound or DMSO for 24 hr in VCaP cells.

Article Snippet: 14 μg of PC3 nuclear extract (Santa Cruz Biotechnology) was added and incubated for 2 hrs at 4 degrees.

Techniques: Biomarker Discovery, Purification, Staining, Western Blot, Viability Assay, Cell Migration Assay, Growth Assay, Migration, Luciferase, Expressing, Control

Figure 3. Effect of TGF-ß signaling upon MED15 expression. qRT-PCR of TGF-b target gene PAI (a) and MED15 (c) of PC3 cells treated with recombinant TGF-b3 for 24 hr. Western blot of p-SMAD3 (b) and MED15 following treatment with recombinant TGF-b3 of PC3 cells for 72 hr (d). Immunohistochemistry of MED15 following treatment with recombinant TGF-b3 in PC3 cells for 72 hr (e,f). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Journal: International journal of cancer

Article Title: MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration-resistant prostate cancer.

doi: 10.1002/ijc.28647

Figure Lengend Snippet: Figure 3. Effect of TGF-ß signaling upon MED15 expression. qRT-PCR of TGF-b target gene PAI (a) and MED15 (c) of PC3 cells treated with recombinant TGF-b3 for 24 hr. Western blot of p-SMAD3 (b) and MED15 following treatment with recombinant TGF-b3 of PC3 cells for 72 hr (d). Immunohistochemistry of MED15 following treatment with recombinant TGF-b3 in PC3 cells for 72 hr (e,f). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Cell lysates and nuclear extracts of PC3 (sc-2152, Santa Cruz, CA) and DU145 (sc-24960, Santa Cruz, CA) were analyzed by immunobloting using MED15 rabbit polyclonal antibody (Proteintech, Chicago, IL).

Techniques: Expressing, Quantitative RT-PCR, Recombinant, Western Blot, Immunohistochemistry

Figure 4. Effect of MED15 on TGF-b signaling and TGF-b-enhanced proliferation. Immunohistochemistry of p-SMAD3 in untreated (a) PC3 cells and following treatment with recombinant TGF-b3 of PC3 cells treated before with scrambled siRNA (b) or siRNA speciﬁc for MED15 (c), as well as Western blotting (d). MTT proliferation assay of untreated and TGF-b3 treated PC3 cells for 72 hr with or without MED15 knockdown. Absorbance of untreated cells was normalized to one. (e). Relative enhancement of absorbance after TGF-b treatment in control and MED15 knockdown PC3 cells (f) *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Journal: International journal of cancer

Article Title: MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration-resistant prostate cancer.

doi: 10.1002/ijc.28647

Figure Lengend Snippet: Figure 4. Effect of MED15 on TGF-b signaling and TGF-b-enhanced proliferation. Immunohistochemistry of p-SMAD3 in untreated (a) PC3 cells and following treatment with recombinant TGF-b3 of PC3 cells treated before with scrambled siRNA (b) or siRNA speciﬁc for MED15 (c), as well as Western blotting (d). MTT proliferation assay of untreated and TGF-b3 treated PC3 cells for 72 hr with or without MED15 knockdown. Absorbance of untreated cells was normalized to one. (e). Relative enhancement of absorbance after TGF-b treatment in control and MED15 knockdown PC3 cells (f) *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Techniques: Immunohistochemistry, Recombinant, Western Blot, Proliferation Assay, Knockdown, Control

Figure 5. MED15 knockdown and proliferation in PCa cells. Correlation of MED15 expression with tumor proliferation markers Ki67 and PHH3 (a). Expression of MED15 in PC3 (b) and DU145 (c) nuclear extracts by Western blot. SiRNA mediated MED15 knockdown in DU145 and PC3 cell lines by Western blot (d) and IHC (e). MTT proliferation assay of PC3 cells (f). *p < 0.05, **p < 0.01, ***p < 0.001. Similarly, proliferation assay of DU145 cells (g). *p < 0.05, **p < 0.01, ***p < 0.001. SiRNA mediated MED15 knockdown in LNCaP cell line (h). Prolif- eration assay of LNCaP cells treated with DHT (i). *p < 0.05, **p < 0.01, ***p < 0.001. ***p < 0.001, Pearson. ShRNA stable knockdown of MED15 in the DU145 cell line (j). Proliferation assay of DU145 cells with stable knockdown of MED15 compared to the scrambled control (k). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Journal: International journal of cancer

Article Title: MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration-resistant prostate cancer.

doi: 10.1002/ijc.28647

Figure Lengend Snippet: Figure 5. MED15 knockdown and proliferation in PCa cells. Correlation of MED15 expression with tumor proliferation markers Ki67 and PHH3 (a). Expression of MED15 in PC3 (b) and DU145 (c) nuclear extracts by Western blot. SiRNA mediated MED15 knockdown in DU145 and PC3 cell lines by Western blot (d) and IHC (e). MTT proliferation assay of PC3 cells (f). *p < 0.05, **p < 0.01, ***p < 0.001. Similarly, proliferation assay of DU145 cells (g). *p < 0.05, **p < 0.01, ***p < 0.001. SiRNA mediated MED15 knockdown in LNCaP cell line (h). Prolif- eration assay of LNCaP cells treated with DHT (i). *p < 0.05, **p < 0.01, ***p < 0.001. ***p < 0.001, Pearson. ShRNA stable knockdown of MED15 in the DU145 cell line (j). Proliferation assay of DU145 cells with stable knockdown of MED15 compared to the scrambled control (k). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color ﬁgure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Techniques: Knockdown, Expressing, Western Blot, Proliferation Assay, shRNA, Control

Expression of ERG, ETS-1, ETS-2 and ETV-4 in PC3 cell nuclear extracts and PCa tissues. Western blot analysis showed that ERG, ETS-1, ETS-2 and ETV-4 were expressed in (A) PC3 cell nuclear extracts, as well as in protein lysates prepared from (B) the PCa tissue samples of five patients.

Journal: International Journal of Molecular Medicine

Article Title: ERG is specifically associated with ETS-2 and ETV-4, but not with ETS-1, in prostate cancer

doi: 10.3892/ijmm.2012.1097

Figure Lengend Snippet: Expression of ERG, ETS-1, ETS-2 and ETV-4 in PC3 cell nuclear extracts and PCa tissues. Western blot analysis showed that ERG, ETS-1, ETS-2 and ETV-4 were expressed in (A) PC3 cell nuclear extracts, as well as in protein lysates prepared from (B) the PCa tissue samples of five patients.

Article Snippet: To investigate whether ERG is associated with ETS-1, ETS-2 and ETV-4 in PC3 cell nuclear extracts (Santa Cruz Biotechnology, Inc.), we performed IP using a rabbit polyclonal anti-ERG antibody (Gentex, Zeeland, MI, USA) as it exhibited the best compatibility with our IP compared to all the other commercially available antibodies that we tested.

Techniques: Expressing, Western Blot

ERG is specifically associated with ETS-2 and ETV-4, but not with ETS-1, in PC3 cell nuclear extracts. Immunoprecipitation (IP) using an anti-ERG antibody showed that ERG is specifically associated with (A) ETS-2 and ETV-4, but not with (B) ETS-1, in PC3 cell nuclear extracts.

Journal: International Journal of Molecular Medicine

Article Title: ERG is specifically associated with ETS-2 and ETV-4, but not with ETS-1, in prostate cancer

doi: 10.3892/ijmm.2012.1097

Figure Lengend Snippet: ERG is specifically associated with ETS-2 and ETV-4, but not with ETS-1, in PC3 cell nuclear extracts. Immunoprecipitation (IP) using an anti-ERG antibody showed that ERG is specifically associated with (A) ETS-2 and ETV-4, but not with (B) ETS-1, in PC3 cell nuclear extracts.

Techniques: Immunoprecipitation

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pc3 nuclear extract (Santa Cruz Biotechnology)

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