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Cell Signaling Technology Inc rabbit anti sp1
siRNA -mediated depletion of <t>SP1</t> or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P
Rabbit Anti Sp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Hypomethylation associated enhanced transcription of trefoil factor-3 mediates tamoxifen-stimulated oncogenicity of ER+ endometrial carcinoma cells"

Article Title: Hypomethylation associated enhanced transcription of trefoil factor-3 mediates tamoxifen-stimulated oncogenicity of ER+ endometrial carcinoma cells

Journal: Oncotarget

doi: 10.18632/oncotarget.20461

siRNA -mediated depletion of SP1 or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P
Figure Legend Snippet: siRNA -mediated depletion of SP1 or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P

Techniques Used: Expressing, Activity Assay, Cell Culture, Transfection, Luciferase, Two Tailed Test

TAM stimulates hypomethylation in the TFF3 promoter and modulates TFF3 transcription through c-JUN/SP1 (A) TFF3 promoter sequence (from -700 to +50bp) located on chromosome 21q22.3 (21:42316053-42314804). Thirteen CpG islands are positioned in the TFF3 promoter sequence, highlighted in red colour. Four binding sites for transcription factors, FOXA2 ( TATTTGATTTTA , -667 to -656bp), c-JUN ( TGTTTCA , -532 to -525bp; TGACTCA , -498 to -491bp), SP1 ( CACCACACCC , -379 to -369bp), and SOX5 ( CAAACAATCC , -119 to -110bp) were identified in the TFF3 promoter sequence, highlighted in bold font. Starting site, ATG, highlighted in green colour. Primer sites highlighted in black (P1) and grey (P2) colour. (B) PCR for bisulfite treated DNA . Total DNA was extracted from cells and treated with bisulfite as described in materials and methods. Sequence of unmethylated (U) and methylated (M) specific primers described in materials and methods. Primer specific sequence amplified on TFF3 promoter mentioned left side . The sizes of detected amplified product in base pair (bp) are shown on the right side. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. (C) TFF3 promoter activity in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. The luciferase assay was performed as described in Materials and Methods. NT , negative control ( pGL3 basic construct ); PT , positive control ( pGL3-TFF3 construct ); pGL3-TFF3 construct with altered sequence of FOXA, c-JUN, SP1 , or SOX5 binding site (mentioned in Figure 2A ), respectively. Construct information is described in methodology section. (D) ChIP analysis in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. ChIP assay was carried out using IgG (control) or SP1 antibody and c-JUN antibody and binding of SP1 and c-JUN was assessed using q-PCR as described in the methodology section. For each PCR, enrichment represent relative to the percentage of Input, error bars represent ±SD . FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test . Columns are mean of triplicate experiments; bars, ± SD . ** P
Figure Legend Snippet: TAM stimulates hypomethylation in the TFF3 promoter and modulates TFF3 transcription through c-JUN/SP1 (A) TFF3 promoter sequence (from -700 to +50bp) located on chromosome 21q22.3 (21:42316053-42314804). Thirteen CpG islands are positioned in the TFF3 promoter sequence, highlighted in red colour. Four binding sites for transcription factors, FOXA2 ( TATTTGATTTTA , -667 to -656bp), c-JUN ( TGTTTCA , -532 to -525bp; TGACTCA , -498 to -491bp), SP1 ( CACCACACCC , -379 to -369bp), and SOX5 ( CAAACAATCC , -119 to -110bp) were identified in the TFF3 promoter sequence, highlighted in bold font. Starting site, ATG, highlighted in green colour. Primer sites highlighted in black (P1) and grey (P2) colour. (B) PCR for bisulfite treated DNA . Total DNA was extracted from cells and treated with bisulfite as described in materials and methods. Sequence of unmethylated (U) and methylated (M) specific primers described in materials and methods. Primer specific sequence amplified on TFF3 promoter mentioned left side . The sizes of detected amplified product in base pair (bp) are shown on the right side. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. (C) TFF3 promoter activity in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. The luciferase assay was performed as described in Materials and Methods. NT , negative control ( pGL3 basic construct ); PT , positive control ( pGL3-TFF3 construct ); pGL3-TFF3 construct with altered sequence of FOXA, c-JUN, SP1 , or SOX5 binding site (mentioned in Figure 2A ), respectively. Construct information is described in methodology section. (D) ChIP analysis in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. ChIP assay was carried out using IgG (control) or SP1 antibody and c-JUN antibody and binding of SP1 and c-JUN was assessed using q-PCR as described in the methodology section. For each PCR, enrichment represent relative to the percentage of Input, error bars represent ±SD . FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test . Columns are mean of triplicate experiments; bars, ± SD . ** P

Techniques Used: Sequencing, Binding Assay, Polymerase Chain Reaction, Methylation, Amplification, Cell Culture, Activity Assay, Luciferase, Negative Control, Construct, Positive Control, Chromatin Immunoprecipitation, Two Tailed Test

2) Product Images from "Lineage-Specific Functionality of an Interferon Regulatory Factor 5 Lupus Risk Haplotype: Lack of B Cell Intrinsic Effects"

Article Title: Lineage-Specific Functionality of an Interferon Regulatory Factor 5 Lupus Risk Haplotype: Lack of B Cell Intrinsic Effects

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00996

The interferon regulatory factor 5 (IRF5) promoter risk insertion/deletion (indel) results in increased IRF5 transcription and specificity protein 1 (SP1) binding in myeloid cells. (A) Normalized activities of non-risk and risk indel promoter-driven luciferase transcription in various cell lines (left), fold change in luciferase activity due to the risk indel in various cell lines (center), and risk, non-risk, and mutated promoter-driven luciferase transcription in THP-1 cells (right). Results of three or four independent experiments are shown. (B) Electrophoretic mobility shift assay showing binding to the 3× and 4× CGGGG indel by various nuclear extracts (left), supershift assay showing SP1 binding to the 3× and 4× CGGGG indel in nuclear extracts of THP-1 cells (right). Results of three independent experiments are shown. (C) SP1 mRNA levels in various cell lines. Results of three independent experiments are shown. p
Figure Legend Snippet: The interferon regulatory factor 5 (IRF5) promoter risk insertion/deletion (indel) results in increased IRF5 transcription and specificity protein 1 (SP1) binding in myeloid cells. (A) Normalized activities of non-risk and risk indel promoter-driven luciferase transcription in various cell lines (left), fold change in luciferase activity due to the risk indel in various cell lines (center), and risk, non-risk, and mutated promoter-driven luciferase transcription in THP-1 cells (right). Results of three or four independent experiments are shown. (B) Electrophoretic mobility shift assay showing binding to the 3× and 4× CGGGG indel by various nuclear extracts (left), supershift assay showing SP1 binding to the 3× and 4× CGGGG indel in nuclear extracts of THP-1 cells (right). Results of three independent experiments are shown. (C) SP1 mRNA levels in various cell lines. Results of three independent experiments are shown. p

Techniques Used: Binding Assay, Luciferase, Activity Assay, Electrophoretic Mobility Shift Assay

3) Product Images from "Lineage-Specific Functionality of an Interferon Regulatory Factor 5 Lupus Risk Haplotype: Lack of B Cell Intrinsic Effects"

Article Title: Lineage-Specific Functionality of an Interferon Regulatory Factor 5 Lupus Risk Haplotype: Lack of B Cell Intrinsic Effects

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00996

The interferon regulatory factor 5 (IRF5) promoter risk insertion/deletion (indel) results in increased IRF5 transcription and specificity protein 1 (SP1) binding in myeloid cells. (A) Normalized activities of non-risk and risk indel promoter-driven luciferase transcription in various cell lines (left), fold change in luciferase activity due to the risk indel in various cell lines (center), and risk, non-risk, and mutated promoter-driven luciferase transcription in THP-1 cells (right). Results of three or four independent experiments are shown. (B) Electrophoretic mobility shift assay showing binding to the 3× and 4× CGGGG indel by various nuclear extracts (left), supershift assay showing SP1 binding to the 3× and 4× CGGGG indel in nuclear extracts of THP-1 cells (right). Results of three independent experiments are shown. (C) SP1 mRNA levels in various cell lines. Results of three independent experiments are shown. p
Figure Legend Snippet: The interferon regulatory factor 5 (IRF5) promoter risk insertion/deletion (indel) results in increased IRF5 transcription and specificity protein 1 (SP1) binding in myeloid cells. (A) Normalized activities of non-risk and risk indel promoter-driven luciferase transcription in various cell lines (left), fold change in luciferase activity due to the risk indel in various cell lines (center), and risk, non-risk, and mutated promoter-driven luciferase transcription in THP-1 cells (right). Results of three or four independent experiments are shown. (B) Electrophoretic mobility shift assay showing binding to the 3× and 4× CGGGG indel by various nuclear extracts (left), supershift assay showing SP1 binding to the 3× and 4× CGGGG indel in nuclear extracts of THP-1 cells (right). Results of three independent experiments are shown. (C) SP1 mRNA levels in various cell lines. Results of three independent experiments are shown. p

Techniques Used: Binding Assay, Luciferase, Activity Assay, Electrophoretic Mobility Shift Assay

4) Product Images from "Peretinoin, an acyclic retinoid, inhibits hepatocarcinogenesis by suppressing sphingosine kinase 1 expression in vitro and in vivo"

Article Title: Peretinoin, an acyclic retinoid, inhibits hepatocarcinogenesis by suppressing sphingosine kinase 1 expression in vitro and in vivo

Journal: Scientific Reports

doi: 10.1038/s41598-017-17285-2

Effects of peretinoin on SPHK1 promoter activity. A effects of peretinoin on the promoter activity of SPHK1 and SPHK2. ( a ) Plasmids encoding luciferase under the control of the promoter region of SPHK1, SPHK2, β-actin, and GAPDH were transfected into Huh-7 cells and, 24 h later, DMSO or peretinoin at 1, 5, 10, 20, or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. The luciferase activity in each condition was normalized to that of DMSO control. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by one-way ANOVA. ( b ) Possible binding sites of Sp1 in the SPHK1 promoter region and a representation of deletion mutants. This figure shows the possible binding sites of Sp1 in the promoter region of SPHK1 and the series of deletion mutants analyzed. ( c ) Enhancement of the promoter activity of SPHK1 by Sp1 overexpression. A plasmid encoding Sp1 or a control empty vector was co-transfected with a plasmid encoding luciferase under the control of the SPHK1 promoter region and, 48 h later, luciferase activity was determined. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by the Student’s t test. ( d ) Restoration of the suppressive effects of peretinoin on the promoter activity of SPHK1 by Sp1 overexpression. A plasmid encoding Sp1 or a control empty vector was transfected into Huh-7 cells. After 24 h, the cells were transfected with a plasmid encoding luciferase under the control of the promoter region of SPHK1 or a control vector without any promoter regions. After another 24 h, DMSO or peretinoin at 20 or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity from peretinoin-treated cells to that of DMSO-treated cells in cells transfected with the plasmid encoding Sp1 or the control empty vector. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by the Student’s t test. ( e ), attenuation of the suppressive effects of peretinoin on the promoter activity of SPHK1 by Sp1 knockdown. Three kinds of siRNA to Sp1, control siRNA (siCNT), or mock were transfected at 20 nM into Huh-7 cells and, 48 h later, the cells were transfected with the plasmid encoding luciferase under the control of the promoter region of SPHK1 or a control vector without any promoter regions. After another 24 h, DMSO or peretinoin at 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity of cells treated with 40 μM peretinoin to that of DMSO-treated cells for each siRNA or mock transfection. ( f ), effects of deletion of the Sp1 binding site on the promoter activity of SPHK1. The plasmids encoding luciferase under the control of various lengths of promoter regions of SPHK1 were transfected into Huh-7 cells and, 24 h later, DMSO or peretinoin at 20 or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity of peretinoin-treated cells to that of DMSO-treated cells for each plasmid. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by two-way ANOVA. *p
Figure Legend Snippet: Effects of peretinoin on SPHK1 promoter activity. A effects of peretinoin on the promoter activity of SPHK1 and SPHK2. ( a ) Plasmids encoding luciferase under the control of the promoter region of SPHK1, SPHK2, β-actin, and GAPDH were transfected into Huh-7 cells and, 24 h later, DMSO or peretinoin at 1, 5, 10, 20, or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. The luciferase activity in each condition was normalized to that of DMSO control. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by one-way ANOVA. ( b ) Possible binding sites of Sp1 in the SPHK1 promoter region and a representation of deletion mutants. This figure shows the possible binding sites of Sp1 in the promoter region of SPHK1 and the series of deletion mutants analyzed. ( c ) Enhancement of the promoter activity of SPHK1 by Sp1 overexpression. A plasmid encoding Sp1 or a control empty vector was co-transfected with a plasmid encoding luciferase under the control of the SPHK1 promoter region and, 48 h later, luciferase activity was determined. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by the Student’s t test. ( d ) Restoration of the suppressive effects of peretinoin on the promoter activity of SPHK1 by Sp1 overexpression. A plasmid encoding Sp1 or a control empty vector was transfected into Huh-7 cells. After 24 h, the cells were transfected with a plasmid encoding luciferase under the control of the promoter region of SPHK1 or a control vector without any promoter regions. After another 24 h, DMSO or peretinoin at 20 or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity from peretinoin-treated cells to that of DMSO-treated cells in cells transfected with the plasmid encoding Sp1 or the control empty vector. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by the Student’s t test. ( e ), attenuation of the suppressive effects of peretinoin on the promoter activity of SPHK1 by Sp1 knockdown. Three kinds of siRNA to Sp1, control siRNA (siCNT), or mock were transfected at 20 nM into Huh-7 cells and, 48 h later, the cells were transfected with the plasmid encoding luciferase under the control of the promoter region of SPHK1 or a control vector without any promoter regions. After another 24 h, DMSO or peretinoin at 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity of cells treated with 40 μM peretinoin to that of DMSO-treated cells for each siRNA or mock transfection. ( f ), effects of deletion of the Sp1 binding site on the promoter activity of SPHK1. The plasmids encoding luciferase under the control of various lengths of promoter regions of SPHK1 were transfected into Huh-7 cells and, 24 h later, DMSO or peretinoin at 20 or 40 μM was added. After 48-h DMSO or peretinoin treatment, luciferase activity was determined. This figure shows the relative ratio of the luciferase activity of peretinoin-treated cells to that of DMSO-treated cells for each plasmid. Error bars indicate the standard deviation from three experiments and the statistical significance of the difference in the average was analyzed by two-way ANOVA. *p

Techniques Used: Activity Assay, Luciferase, Transfection, Standard Deviation, Binding Assay, Over Expression, Plasmid Preparation

Related Articles

Incubation:

Article Title: MiR-150-3p targets SP1 and suppresses the growth of glioma cells
Article Snippet: .. The protein was then transferred on to the nitrocellulose membrane (Millipore, Billerica, MA, U.S.A.) and blocked with 5% of non-fat milk at RT for 1 h. Subsequently, membrane was incubated with primary antibodies against PTEN (ab32199, 1:2000 dilution, Abcam), GAPDH (sc-20357, 1:3000 dilution, Santa Cruz Biotechnology, Inc., Dallas, TX, U.S.A.), SP1 (#5931, 1:2000 dilution, Cell Signaling Technology, Danvers, MA, U.S.A.) for 2 h at RT, respectively. .. After washing with TBS and Tween-20 (TBST), the membrane was incubated with the secondary antibody conjugated with horseradish peroxidase (HRP) for 1 h at RT.

Article Title: The Sp1 transcription factor is essential for the expression of gliostatin/thymidine phosphorylase in rheumatoid fibroblast-like synoviocytes
Article Snippet: .. The blots were blocked for 60 minutes with 5% non-fat dry milk in Tris-buffered saline containing 0.1% Tween 20 (TBS-T) and then incubated with an anti-Sp1 antibody (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-lamin C (1:1,000; ImmuQuest Ltd, Seamer, North Yorkshire, UK), and anti-α-tubulin antibody (1:1,000; Cell Signaling Technology, Inc.) as loading controls in TBS-T for 2 hours at room temperature. .. After three washes with TBS-T, membranes were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies in TBS-T (1:3,000; GE Healthcare, Little Chalfont, Buckinghamshire, UK) for 1 hour at room temperature and washed three times with TBS-T.

Article Title: Nicotine stimulates PPAR?/? expression in human lung carcinoma cells through activation of PI3-K/mTOR and suppression of AP-2?
Article Snippet: .. Blots were incubated with primary antibodies against PPARβ/δ, α7 nAChR, AP-2α, AP-2β, AP-2γ or Sp1 (1:1000) overnight at 4°C, washed, and incubated with secondary anti-rabbit IgG conjugated to horseradish peroxidase (1:2,000 dilution, Cell Signaling) for 1 h at room temperature. .. Blots were transferred to ECL solution (Pierce, Rockford IL), exposed to X-ray film and proteins were quantified by densitometric scanning using a Bio-Rad GS-800 calibrated densitometer.

Chromatin Immunoprecipitation:

Article Title: The Cyclin-dependent Kinase Inhibitor p16INK4a Physically Interacts with Transcription Factor Sp1 and Cyclin-dependent Kinase 4 to Transactivate MicroRNA-141 and MicroRNA-146b-5p Spontaneously and in Response to Ultraviolet Light-induced DNA Damage *
Article Snippet: .. ChIP experiments were performed using antibodies against Sp1 (Cell Signaling Technology), p16 (BD Biosciences), and CDK4 (C-22). ..

Blocking Assay:

Article Title: Hyperglycemia and hyperlipidemia blunts the Insulin-Inpp5f negative feedback loop in the diabetic heart
Article Snippet: .. After blocking with 5% non-fat milk, the membranes were probed overnight at 4 °C with Inpp5f (1:500, cat# SAB2700848, Sigma, USA), p-Akt Ser473 (1:1000, cat# 9271, Cell signaling, USA), total Akt (1:1000, cat# 9272, Cell signaling, USA), p65 (1:1000, ab32536, Abcam, USA), Sp1 (1:1000, cat# 5931, Cell signaling, USA), β-actin (1:1000, cat# AB10024, Sangon, China), Lamin B (1:500, cat# BA1228, Boster, China) antibodies. .. Secondary antibodies conjugated to IRDye TM 800 (1:15 000, Rockland, USA) were detected using an Odyssey infrared imaging system (LI-COR, USA).

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    Cell Signaling Technology Inc rabbit anti sp1
    siRNA -mediated depletion of <t>SP1</t> or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P
    Rabbit Anti Sp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti sp1/product/Cell Signaling Technology Inc
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti sp1 - by Bioz Stars, 2020-08
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    Cell Signaling Technology Inc rabbit monoclonal ab against sp1
    ZEB1 activates VEGFA transcription by recruiting <t>SP1</t> to the endogenous VEGFA promoter. (A) Sequential deletion and mutation of SP1 elements on the human VEGFA promoter were fused to the luciferase reporter. (B) MDA-MB-231 cells were co-transfected with the ZEB1 expression plasmid (1 μg/well) and different wild-type VEGFA promoter luciferase reporter constructs (1 μg/well). Extract luciferase activities were determined 48 h after transfection using a Betascope analyzer. Luciferase values were normalized to Renilla activities. * P
    Rabbit Monoclonal Ab Against Sp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal ab against sp1/product/Cell Signaling Technology Inc
    Average 90 stars, based on 1 article reviews
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    94
    Cell Signaling Technology Inc rabbit anti sp1 monoclonal antibody
    Western blot analysis of <t>Sp1</t> protein in 2DG-treated NCCIT cells. The whole cell lysates of NCCIT cells treated with 2DG for the indicated times (0, 24, 72, or 168 h) were precipitated with an anti-Sp1 antibody (D4C3), and the precipitated proteins were immunoblotted with D4C3, anti- O -GlcNAc antibodies (RL2 or CTD110.6), anti-Phosphoserine/threonine (P-Ser/Thr), anti-SUMO1, or anti-Ubiquitin. These transcriptional modifications were observed in Sp1 proteins [6] . Among these modifications, only O -GlcNAcylation was clearly detected in the D4C3 precipitates. The Sp1 levels in whole cell lysates were shown as a reference (left panel).
    Rabbit Anti Sp1 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti sp1 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Cell Signaling Technology Inc rabbit anti human sp1 antibody
    <t>SP1</t> was a directly targeted gene of miR-149 in vitro . (A) Bioinformatics software assessed that SP1 mRNA contained a miR-149 seed match at position 3809–3815 of the SP1 3′-UTR. (B) miR-149 significantly inhibited the SP1 Wt but not the
    Rabbit Anti Human Sp1 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    siRNA -mediated depletion of SP1 or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P

    Journal: Oncotarget

    Article Title: Hypomethylation associated enhanced transcription of trefoil factor-3 mediates tamoxifen-stimulated oncogenicity of ER+ endometrial carcinoma cells

    doi: 10.18632/oncotarget.20461

    Figure Lengend Snippet: siRNA -mediated depletion of SP1 or c-JUN expression in ER+ EC cells abolished TAM-stimulated promoter activity of TFF3 and growth in Matrigel culture (A) TFF3 promoter activity and (B) growth in Matrigel of EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. Depletion of SP1 or c-JUN expression was achieved using transient-transfection of si - RNA directed to SP1 or c-JUN transcript respectively, as described in materials and methods. Universal controls (scrambled oligo) were used for transfection control as described in materials and methods. The luciferase assay was performed as described in materials and methods. FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test ( P

    Article Snippet: IHC analysis was performed as previously described [ ] using rabbit anti-TFF3 was obtained from Abcam, Cambridge, MA; rabbit anti-SP1, rabbit anti-p-c-JUN and rabbit anti-c-JUN antibodies wereobtained from Cell Signaling Technology, Singapore [ ].

    Techniques: Expressing, Activity Assay, Cell Culture, Transfection, Luciferase, Two Tailed Test

    TAM stimulates hypomethylation in the TFF3 promoter and modulates TFF3 transcription through c-JUN/SP1 (A) TFF3 promoter sequence (from -700 to +50bp) located on chromosome 21q22.3 (21:42316053-42314804). Thirteen CpG islands are positioned in the TFF3 promoter sequence, highlighted in red colour. Four binding sites for transcription factors, FOXA2 ( TATTTGATTTTA , -667 to -656bp), c-JUN ( TGTTTCA , -532 to -525bp; TGACTCA , -498 to -491bp), SP1 ( CACCACACCC , -379 to -369bp), and SOX5 ( CAAACAATCC , -119 to -110bp) were identified in the TFF3 promoter sequence, highlighted in bold font. Starting site, ATG, highlighted in green colour. Primer sites highlighted in black (P1) and grey (P2) colour. (B) PCR for bisulfite treated DNA . Total DNA was extracted from cells and treated with bisulfite as described in materials and methods. Sequence of unmethylated (U) and methylated (M) specific primers described in materials and methods. Primer specific sequence amplified on TFF3 promoter mentioned left side . The sizes of detected amplified product in base pair (bp) are shown on the right side. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. (C) TFF3 promoter activity in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. The luciferase assay was performed as described in Materials and Methods. NT , negative control ( pGL3 basic construct ); PT , positive control ( pGL3-TFF3 construct ); pGL3-TFF3 construct with altered sequence of FOXA, c-JUN, SP1 , or SOX5 binding site (mentioned in Figure 2A ), respectively. Construct information is described in methodology section. (D) ChIP analysis in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. ChIP assay was carried out using IgG (control) or SP1 antibody and c-JUN antibody and binding of SP1 and c-JUN was assessed using q-PCR as described in the methodology section. For each PCR, enrichment represent relative to the percentage of Input, error bars represent ±SD . FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test . Columns are mean of triplicate experiments; bars, ± SD . ** P

    Journal: Oncotarget

    Article Title: Hypomethylation associated enhanced transcription of trefoil factor-3 mediates tamoxifen-stimulated oncogenicity of ER+ endometrial carcinoma cells

    doi: 10.18632/oncotarget.20461

    Figure Lengend Snippet: TAM stimulates hypomethylation in the TFF3 promoter and modulates TFF3 transcription through c-JUN/SP1 (A) TFF3 promoter sequence (from -700 to +50bp) located on chromosome 21q22.3 (21:42316053-42314804). Thirteen CpG islands are positioned in the TFF3 promoter sequence, highlighted in red colour. Four binding sites for transcription factors, FOXA2 ( TATTTGATTTTA , -667 to -656bp), c-JUN ( TGTTTCA , -532 to -525bp; TGACTCA , -498 to -491bp), SP1 ( CACCACACCC , -379 to -369bp), and SOX5 ( CAAACAATCC , -119 to -110bp) were identified in the TFF3 promoter sequence, highlighted in bold font. Starting site, ATG, highlighted in green colour. Primer sites highlighted in black (P1) and grey (P2) colour. (B) PCR for bisulfite treated DNA . Total DNA was extracted from cells and treated with bisulfite as described in materials and methods. Sequence of unmethylated (U) and methylated (M) specific primers described in materials and methods. Primer specific sequence amplified on TFF3 promoter mentioned left side . The sizes of detected amplified product in base pair (bp) are shown on the right side. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. (C) TFF3 promoter activity in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. The luciferase assay was performed as described in Materials and Methods. NT , negative control ( pGL3 basic construct ); PT , positive control ( pGL3-TFF3 construct ); pGL3-TFF3 construct with altered sequence of FOXA, c-JUN, SP1 , or SOX5 binding site (mentioned in Figure 2A ), respectively. Construct information is described in methodology section. (D) ChIP analysis in EC cells. EC cells (VE or CTE) were cultured in FM or CSF-PRFM. 5μM TAM was used to treat cells. ChIP assay was carried out using IgG (control) or SP1 antibody and c-JUN antibody and binding of SP1 and c-JUN was assessed using q-PCR as described in the methodology section. For each PCR, enrichment represent relative to the percentage of Input, error bars represent ±SD . FM were 10%FBS, standard media conditions as per ATCC propagation instructions; and CSF-PRFM were charcoal striped 10% FBS, phenol-red free media. Statistical significance was assessed by using an unpaired two-tailed Student's t test . Columns are mean of triplicate experiments; bars, ± SD . ** P

    Article Snippet: IHC analysis was performed as previously described [ ] using rabbit anti-TFF3 was obtained from Abcam, Cambridge, MA; rabbit anti-SP1, rabbit anti-p-c-JUN and rabbit anti-c-JUN antibodies wereobtained from Cell Signaling Technology, Singapore [ ].

    Techniques: Sequencing, Binding Assay, Polymerase Chain Reaction, Methylation, Amplification, Cell Culture, Activity Assay, Luciferase, Negative Control, Construct, Positive Control, Chromatin Immunoprecipitation, Two Tailed Test

    ZEB1 activates VEGFA transcription by recruiting SP1 to the endogenous VEGFA promoter. (A) Sequential deletion and mutation of SP1 elements on the human VEGFA promoter were fused to the luciferase reporter. (B) MDA-MB-231 cells were co-transfected with the ZEB1 expression plasmid (1 μg/well) and different wild-type VEGFA promoter luciferase reporter constructs (1 μg/well). Extract luciferase activities were determined 48 h after transfection using a Betascope analyzer. Luciferase values were normalized to Renilla activities. * P

    Journal: PLoS ONE

    Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

    doi: 10.1371/journal.pone.0148774

    Figure Lengend Snippet: ZEB1 activates VEGFA transcription by recruiting SP1 to the endogenous VEGFA promoter. (A) Sequential deletion and mutation of SP1 elements on the human VEGFA promoter were fused to the luciferase reporter. (B) MDA-MB-231 cells were co-transfected with the ZEB1 expression plasmid (1 μg/well) and different wild-type VEGFA promoter luciferase reporter constructs (1 μg/well). Extract luciferase activities were determined 48 h after transfection using a Betascope analyzer. Luciferase values were normalized to Renilla activities. * P

    Article Snippet: The Abs used in these experiments were rabbit monoclonal Ab against SP1 (#9389S; CST) and anti-rabbit normal IgG (sc-2345, Santa Cruz).

    Techniques: Mutagenesis, Luciferase, Multiple Displacement Amplification, Transfection, Expressing, Plasmid Preparation, Construct

    Western blot analysis of Sp1 protein in 2DG-treated NCCIT cells. The whole cell lysates of NCCIT cells treated with 2DG for the indicated times (0, 24, 72, or 168 h) were precipitated with an anti-Sp1 antibody (D4C3), and the precipitated proteins were immunoblotted with D4C3, anti- O -GlcNAc antibodies (RL2 or CTD110.6), anti-Phosphoserine/threonine (P-Ser/Thr), anti-SUMO1, or anti-Ubiquitin. These transcriptional modifications were observed in Sp1 proteins [6] . Among these modifications, only O -GlcNAcylation was clearly detected in the D4C3 precipitates. The Sp1 levels in whole cell lysates were shown as a reference (left panel).

    Journal: Data in Brief

    Article Title: Western blot data using two distinct anti-O-GlcNAc monoclonal antibodies showing unique glycosylation status on cellular proteins under 2-deoxy-d-glucose treatment

    doi: 10.1016/j.dib.2016.12.001

    Figure Lengend Snippet: Western blot analysis of Sp1 protein in 2DG-treated NCCIT cells. The whole cell lysates of NCCIT cells treated with 2DG for the indicated times (0, 24, 72, or 168 h) were precipitated with an anti-Sp1 antibody (D4C3), and the precipitated proteins were immunoblotted with D4C3, anti- O -GlcNAc antibodies (RL2 or CTD110.6), anti-Phosphoserine/threonine (P-Ser/Thr), anti-SUMO1, or anti-Ubiquitin. These transcriptional modifications were observed in Sp1 proteins [6] . Among these modifications, only O -GlcNAcylation was clearly detected in the D4C3 precipitates. The Sp1 levels in whole cell lysates were shown as a reference (left panel).

    Article Snippet: The primary antibodies used were mouse anti-O -GlcNAc monoclonal antibodies (RL2, Thermo Fisher Scientific, Waltham, MA; CTD110.6, Cell Signaling Technology), rabbit anti-Sp1 monoclonal antibody (D4C3, Cell Signaling Technology), rabbit anti-GRP78/Bip monoclonal antibody (C50B12, Cell Signaling Technology), rabbit anti-GAPDH monoclonal antibody (D16H11, Cell Signaling Technology), rabbit anti-β-Actin polyclonal antibody (#4967, Cell Signaling Technology), rabbit anti-Phosphoserine/threonine polyclonal antibody (ab17464, abcam, Cambridge, UK), mouse anti-SUMO1 monoclonal antibody (21C7, BostonBiochem, Cambridge, MA), and mouse anti-Ubiquitin monoclonal antibody (P4D1, Cell Signaling Technology).

    Techniques: Western Blot

    SP1 was a directly targeted gene of miR-149 in vitro . (A) Bioinformatics software assessed that SP1 mRNA contained a miR-149 seed match at position 3809–3815 of the SP1 3′-UTR. (B) miR-149 significantly inhibited the SP1 Wt but not the

    Journal: Oncology Letters

    Article Title: MicroRNA-149 targets specificity protein 1 to suppress human tongue squamous cell carcinoma cell proliferation and motility

    doi: 10.3892/ol.2016.5527

    Figure Lengend Snippet: SP1 was a directly targeted gene of miR-149 in vitro . (A) Bioinformatics software assessed that SP1 mRNA contained a miR-149 seed match at position 3809–3815 of the SP1 3′-UTR. (B) miR-149 significantly inhibited the SP1 Wt but not the

    Article Snippet: The membranes were blocked with 5% skimmed milk at room temperature for 2 h and incubated with rabbit anti-human SP1 antibody (dilution, 1: 1,000; cat. no., 5931; Cell Signaling Technology, Inc., Danvers, MA, USA) according to the protocol of the manufacturer at 4°C overnight.

    Techniques: In Vitro, Software