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pgl3 basic 180 luciferase reporter plasmid  (Addgene inc)


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    Addgene inc pgl3 basic 180 luciferase reporter plasmid
    Pgl3 Basic 180 Luciferase Reporter Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 195 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgl3+basic+luciferase+reporter+plasmid/pGL3+(-BRE)+Luciferase+(Plasmid+%2345127)/10__1158_slash_0008___5472__can___25___2893-63-26-31
    Average 95 stars, based on 195 article reviews
    pgl3 basic 180 luciferase reporter plasmid - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Tubule-mitophagic secretion of SerpinG1 reprograms macrophages to instruct anti-septic acute kidney injury efficacy of high-dose ascorbate mediated by NRF2 transactivation
    Article Snippet: .. The pGL3-ARE-luc vector was generated by inserting three copies of the antioxidant response element (ARE; 5'-GTGACAAAGCAATCCCGT GACAAAGCAATCCCGTGACAAAGCAATA-3') into pGL3-basic luciferase reporter plasmid. pLV-H1-SGIPZ-shNRF2 was constructed by inserting NRF2 shRNA into lentiviral pLV-H1-SGIPZ vectors (119217, Addgene). .. Lentiviral shRNA targeting Atg7 (TRCN0000092164) was obtained from the TRC library (Merck).

    Article Title: PPARγ induces NEDD4 gene expression to promote autophagy and insulin action.
    Article Snippet: This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.. Please cite this article as doi: 10.1111/febs.15042 This article is protected by copyright.. All rights reserved.

    Article Title: MicroRNA‑218 inhibits the malignant phenotypes of glioma by modulating the TNC/AKT/AP‑1/TGFβ1 feedback signaling loop.
    Article Snippet: .. The 3xAP in pGL3‐Basic luciferase reporter plasmid was purchased from Addgene, Inc. (plasmid no. 40342), which contains three canonical AP‐1 binding sites (TGACTCA) upstream of the luciferase reporter plasmid pGL3‐Basic promoter fragment. .. To assess the 3'‐UTR activity of TNC mRNA modulated by miR‐218, U251 and SHG44 cells were transfected with NC or miR‐218‐mimics in 6‐well plates and subsequently co‐trans‐ fected with pmirGLO‐TNC 3'‐UTR‐WT or pmirGLO‐TNC 3'‐UTR‐MUT, using Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.).

    Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
    Article Snippet: .. The promoter region of human SQSTM1 was amplified by PCR from the U2OS genome and cloned into the pGL3-basic luciferase reporter plasmid (Addgene). ..

    Article Title: YAP induces an oncogenic transcriptional program through TET1-mediated epigenetic remodeling in liver growth and tumorigenesis.
    Article Snippet: 1Department of Physiology, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA.. 2Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.. ✉e-mail: duojia.pan@utsouthwestern.edu The preexisting epigenetic landscape represents a barrier to oncogene-induced cellular transformation and malignancy.

    Article Title: MiT/ TFE factors control ER ‐phagy via transcriptional regulation of FAM 134B
    Article Snippet: .. The promoter region of rat Fam134b from 78211858 to 78211187 (CLEAR 108404, −2708 from TSS position in intron 3) was amplified by PCR from RCS chondrocyte genome and cloned into pGL3‐basic luciferase reporter plasmid (Addgene). ..

    Generated:

    Article Title: Tubule-mitophagic secretion of SerpinG1 reprograms macrophages to instruct anti-septic acute kidney injury efficacy of high-dose ascorbate mediated by NRF2 transactivation
    Article Snippet: .. The pGL3-ARE-luc vector was generated by inserting three copies of the antioxidant response element (ARE; 5'-GTGACAAAGCAATCCCGT GACAAAGCAATCCCGTGACAAAGCAATA-3') into pGL3-basic luciferase reporter plasmid. pLV-H1-SGIPZ-shNRF2 was constructed by inserting NRF2 shRNA into lentiviral pLV-H1-SGIPZ vectors (119217, Addgene). .. Lentiviral shRNA targeting Atg7 (TRCN0000092164) was obtained from the TRC library (Merck).

    Luciferase:

    Article Title: Tubule-mitophagic secretion of SerpinG1 reprograms macrophages to instruct anti-septic acute kidney injury efficacy of high-dose ascorbate mediated by NRF2 transactivation
    Article Snippet: .. The pGL3-ARE-luc vector was generated by inserting three copies of the antioxidant response element (ARE; 5'-GTGACAAAGCAATCCCGT GACAAAGCAATCCCGTGACAAAGCAATA-3') into pGL3-basic luciferase reporter plasmid. pLV-H1-SGIPZ-shNRF2 was constructed by inserting NRF2 shRNA into lentiviral pLV-H1-SGIPZ vectors (119217, Addgene). .. Lentiviral shRNA targeting Atg7 (TRCN0000092164) was obtained from the TRC library (Merck).

    Article Title: PPARγ induces NEDD4 gene expression to promote autophagy and insulin action.
    Article Snippet: This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.. Please cite this article as doi: 10.1111/febs.15042 This article is protected by copyright.. All rights reserved.

    Article Title: MicroRNA‑218 inhibits the malignant phenotypes of glioma by modulating the TNC/AKT/AP‑1/TGFβ1 feedback signaling loop.
    Article Snippet: .. The 3xAP in pGL3‐Basic luciferase reporter plasmid was purchased from Addgene, Inc. (plasmid no. 40342), which contains three canonical AP‐1 binding sites (TGACTCA) upstream of the luciferase reporter plasmid pGL3‐Basic promoter fragment. .. To assess the 3'‐UTR activity of TNC mRNA modulated by miR‐218, U251 and SHG44 cells were transfected with NC or miR‐218‐mimics in 6‐well plates and subsequently co‐trans‐ fected with pmirGLO‐TNC 3'‐UTR‐WT or pmirGLO‐TNC 3'‐UTR‐MUT, using Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.).

    Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
    Article Snippet: .. The promoter region of human SQSTM1 was amplified by PCR from the U2OS genome and cloned into the pGL3-basic luciferase reporter plasmid (Addgene). ..

    Article Title: YAP induces an oncogenic transcriptional program through TET1-mediated epigenetic remodeling in liver growth and tumorigenesis.
    Article Snippet: 1Department of Physiology, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA.. 2Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.. ✉e-mail: duojia.pan@utsouthwestern.edu The preexisting epigenetic landscape represents a barrier to oncogene-induced cellular transformation and malignancy.

    Article Title: MiT/ TFE factors control ER ‐phagy via transcriptional regulation of FAM 134B
    Article Snippet: .. The promoter region of rat Fam134b from 78211858 to 78211187 (CLEAR 108404, −2708 from TSS position in intron 3) was amplified by PCR from RCS chondrocyte genome and cloned into pGL3‐basic luciferase reporter plasmid (Addgene). ..

    Construct:

    Article Title: Tubule-mitophagic secretion of SerpinG1 reprograms macrophages to instruct anti-septic acute kidney injury efficacy of high-dose ascorbate mediated by NRF2 transactivation
    Article Snippet: .. The pGL3-ARE-luc vector was generated by inserting three copies of the antioxidant response element (ARE; 5'-GTGACAAAGCAATCCCGT GACAAAGCAATCCCGTGACAAAGCAATA-3') into pGL3-basic luciferase reporter plasmid. pLV-H1-SGIPZ-shNRF2 was constructed by inserting NRF2 shRNA into lentiviral pLV-H1-SGIPZ vectors (119217, Addgene). .. Lentiviral shRNA targeting Atg7 (TRCN0000092164) was obtained from the TRC library (Merck).

    Article Title: PPARγ induces NEDD4 gene expression to promote autophagy and insulin action.
    Article Snippet: This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.. Please cite this article as doi: 10.1111/febs.15042 This article is protected by copyright.. All rights reserved.

    shRNA:

    Article Title: Tubule-mitophagic secretion of SerpinG1 reprograms macrophages to instruct anti-septic acute kidney injury efficacy of high-dose ascorbate mediated by NRF2 transactivation
    Article Snippet: .. The pGL3-ARE-luc vector was generated by inserting three copies of the antioxidant response element (ARE; 5'-GTGACAAAGCAATCCCGT GACAAAGCAATCCCGTGACAAAGCAATA-3') into pGL3-basic luciferase reporter plasmid. pLV-H1-SGIPZ-shNRF2 was constructed by inserting NRF2 shRNA into lentiviral pLV-H1-SGIPZ vectors (119217, Addgene). .. Lentiviral shRNA targeting Atg7 (TRCN0000092164) was obtained from the TRC library (Merck).

    Polymerase Chain Reaction:

    Article Title: PPARγ induces NEDD4 gene expression to promote autophagy and insulin action.
    Article Snippet: This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.. Please cite this article as doi: 10.1111/febs.15042 This article is protected by copyright.. All rights reserved.

    Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
    Article Snippet: .. The promoter region of human SQSTM1 was amplified by PCR from the U2OS genome and cloned into the pGL3-basic luciferase reporter plasmid (Addgene). ..

    Article Title: MiT/ TFE factors control ER ‐phagy via transcriptional regulation of FAM 134B
    Article Snippet: .. The promoter region of rat Fam134b from 78211858 to 78211187 (CLEAR 108404, −2708 from TSS position in intron 3) was amplified by PCR from RCS chondrocyte genome and cloned into pGL3‐basic luciferase reporter plasmid (Addgene). ..

    Cloning:

    Article Title: PPARγ induces NEDD4 gene expression to promote autophagy and insulin action.
    Article Snippet: This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record.. Please cite this article as doi: 10.1111/febs.15042 This article is protected by copyright.. All rights reserved.

    Binding Assay:

    Article Title: MicroRNA‑218 inhibits the malignant phenotypes of glioma by modulating the TNC/AKT/AP‑1/TGFβ1 feedback signaling loop.
    Article Snippet: .. The 3xAP in pGL3‐Basic luciferase reporter plasmid was purchased from Addgene, Inc. (plasmid no. 40342), which contains three canonical AP‐1 binding sites (TGACTCA) upstream of the luciferase reporter plasmid pGL3‐Basic promoter fragment. .. To assess the 3'‐UTR activity of TNC mRNA modulated by miR‐218, U251 and SHG44 cells were transfected with NC or miR‐218‐mimics in 6‐well plates and subsequently co‐trans‐ fected with pmirGLO‐TNC 3'‐UTR‐WT or pmirGLO‐TNC 3'‐UTR‐MUT, using Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.).

    Amplification:

    Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
    Article Snippet: .. The promoter region of human SQSTM1 was amplified by PCR from the U2OS genome and cloned into the pGL3-basic luciferase reporter plasmid (Addgene). ..

    Article Title: MiT/ TFE factors control ER ‐phagy via transcriptional regulation of FAM 134B
    Article Snippet: .. The promoter region of rat Fam134b from 78211858 to 78211187 (CLEAR 108404, −2708 from TSS position in intron 3) was amplified by PCR from RCS chondrocyte genome and cloned into pGL3‐basic luciferase reporter plasmid (Addgene). ..

    Clone Assay:

    Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
    Article Snippet: .. The promoter region of human SQSTM1 was amplified by PCR from the U2OS genome and cloned into the pGL3-basic luciferase reporter plasmid (Addgene). ..

    Article Title: MiT/ TFE factors control ER ‐phagy via transcriptional regulation of FAM 134B
    Article Snippet: .. The promoter region of rat Fam134b from 78211858 to 78211187 (CLEAR 108404, −2708 from TSS position in intron 3) was amplified by PCR from RCS chondrocyte genome and cloned into pGL3‐basic luciferase reporter plasmid (Addgene). ..

    other:

    Article Title: MicroRNA-218 inhibits the malignant phenotypes of glioma by modulating the TNC/AKT/AP-1/TGFβ1 feedback signaling loop
    Article Snippet: The wild-type (WT) TNC 3′-UTR was amplified from cDNA of U251 cells to construct the luciferase reporter plasmids The TNC 3′-UTR with mutant (MUT) binding site of miR-218 was synthesized by Sangon Biotech Co, Ltd These two fragments were inserted into pre-digested pmirGLO luciferase vector (gifted by Dr Yanke Chen at Xi'an Jiaotong University Health Science Center) to produce the luciferase reporter plasmids, pmirGLO-TNC 3′-UTR-WT and pmirGLO-TNC 3′-UTR-MUT The primer sequences for plasmid constructs are listed in The 3×AP in pGL3-Basic luciferase reporter plasmid was purchased from Addgene, Inc (plasmid no 40342), which contains three canonical AP-1 binding sites (TGACTCA) upstream of the luciferase reporter plasmid pGL3-Basic promoter fragment To assess the 3′-UTR activity of TNC mRNA modulated by miR-218, U251 and SHG44 cells were transfected with NC or miR-218-mimics in 6-well plates and subsequently co-transfected with pmirGLO-TNC 3′-UTR-WT or pmirGLO-TNC 3′-UTR-MUT, using Lipofectamine ® 3000 (Invitrogen; Thermo Fisher Scientific, Inc) To determine transcriptional activity of AP-1 regulated by miR-218 or TGFβ1, U251 and SHG44 cells were treated with TGFβ1 (10 ng/ml; at 37°C for 24 h) or transfected with NC/miR-218-mimics, and were subsequently co-transfected with pRL-TK plasmids and the 3×AP in pGL3-Basic luciferase reporter plasmid Following incubation for 36 h, luciferase activities were detected using a dual-luciferase reporter assay system (Promega Corporation) on an EnSpire Multimode Plate Reader (PerkinElmer, Inc) Firefly luciferase activity was normalized to Renilla luciferase activity All experiments were performed in triplicate



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    Addgene inc luciferase reporter plasmid 3xap1pgl3
    Estrogen promotes HNRNPAB expression via SP1. (A) Immunoblot assessment of HNRNPAB in 2 human normal breast epithelial cell lines and 11 human BC cell lines. (B) CPTAC database analyzed the relative HNRNPAB and ERα protein level in 105 BC patients. P value was determined by Pearson correlation analysis. (C) The relative expression of TFF1 and HNRNPAB mRNAs after stimulated with 10 nM E 2 in estrogen-deprived MCF-7 cells for 48 h analyzed by qRT-PCR. (D) Immunoblot assessment of HNRNPAB expression in MCF-7 with or without 10 nM E 2 in estrogen-deprived MCF-7 cells for 72 h, or in short-term oestrogen deprivation (STED) MCF-7 cells for 7 days and parental cells. (E) The schematic of HNRNPAB promoter region sequence selected from the transcription start site of -2 kb~+100 bp. (F) MCF-7 cells were transfected with <t>pGL3</t> basic reporter vector containing HNRNPAB promoter, and pRL-TK reporter control vector containing luciferase activity, as well as control siRNA or different siRNA sequences of JUN , FOS , FOXA1 and SP1 . The relative fluorescence activity was measured. (G) CPTAC database analyzed the relative HNRNPAB and SP1 protein level in 68 ER+ BC patients. P value was determined by Pearson correlation analysis. (H) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector containing different regions of HNRNPAB promoter. The relative fluorescence activity was measured. (I) The schematic of three predicted conserved SP1 binding sites on the upstream -425 bp~+100 bp regions of the human HNRNPAB gene. Red boxes represent the predicted SP1 binding sites. (J) The DNA regions enriched by SP1 in ChIP assay were verified by PCR. (K) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector or containing “b” or “b” mutant regions. The relative fluorescence activity was measured. (L) The relative expression of SP1 and HNRNPAB mRNAs in MCF-7 cells bearing control or SP1 shRNAs analyzed by qRT-PCR. (M) Immunoblot assessment of SP1 and HNRNPAB expression in MCF-7 cells with or without SP1 shRNAs in the presence or absence of 10 nM E 2 for 48 h. Data was shown as mean ± S.D. from three independent experiments. Unpaired two-tailed Student's t test (C, H, K) and one-way ANOVA followed by Tukey's multiple comparisons test (F, L). ns, P >0.05; ***, P <0.001; ****, P <0.0001.
    Luciferase Reporter Plasmid 3xap1pgl3, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ribobio co p65 pgl3-basic luciferase reporter plasmid
    Estrogen promotes HNRNPAB expression via SP1. (A) Immunoblot assessment of HNRNPAB in 2 human normal breast epithelial cell lines and 11 human BC cell lines. (B) CPTAC database analyzed the relative HNRNPAB and ERα protein level in 105 BC patients. P value was determined by Pearson correlation analysis. (C) The relative expression of TFF1 and HNRNPAB mRNAs after stimulated with 10 nM E 2 in estrogen-deprived MCF-7 cells for 48 h analyzed by qRT-PCR. (D) Immunoblot assessment of HNRNPAB expression in MCF-7 with or without 10 nM E 2 in estrogen-deprived MCF-7 cells for 72 h, or in short-term oestrogen deprivation (STED) MCF-7 cells for 7 days and parental cells. (E) The schematic of HNRNPAB promoter region sequence selected from the transcription start site of -2 kb~+100 bp. (F) MCF-7 cells were transfected with <t>pGL3</t> basic reporter vector containing HNRNPAB promoter, and pRL-TK reporter control vector containing luciferase activity, as well as control siRNA or different siRNA sequences of JUN , FOS , FOXA1 and SP1 . The relative fluorescence activity was measured. (G) CPTAC database analyzed the relative HNRNPAB and SP1 protein level in 68 ER+ BC patients. P value was determined by Pearson correlation analysis. (H) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector containing different regions of HNRNPAB promoter. The relative fluorescence activity was measured. (I) The schematic of three predicted conserved SP1 binding sites on the upstream -425 bp~+100 bp regions of the human HNRNPAB gene. Red boxes represent the predicted SP1 binding sites. (J) The DNA regions enriched by SP1 in ChIP assay were verified by PCR. (K) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector or containing “b” or “b” mutant regions. The relative fluorescence activity was measured. (L) The relative expression of SP1 and HNRNPAB mRNAs in MCF-7 cells bearing control or SP1 shRNAs analyzed by qRT-PCR. (M) Immunoblot assessment of SP1 and HNRNPAB expression in MCF-7 cells with or without SP1 shRNAs in the presence or absence of 10 nM E 2 for 48 h. Data was shown as mean ± S.D. from three independent experiments. Unpaired two-tailed Student's t test (C, H, K) and one-way ANOVA followed by Tukey's multiple comparisons test (F, L). ns, P >0.05; ***, P <0.001; ****, P <0.0001.
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    Estrogen promotes HNRNPAB expression via SP1. (A) Immunoblot assessment of HNRNPAB in 2 human normal breast epithelial cell lines and 11 human BC cell lines. (B) CPTAC database analyzed the relative HNRNPAB and ERα protein level in 105 BC patients. P value was determined by Pearson correlation analysis. (C) The relative expression of TFF1 and HNRNPAB mRNAs after stimulated with 10 nM E 2 in estrogen-deprived MCF-7 cells for 48 h analyzed by qRT-PCR. (D) Immunoblot assessment of HNRNPAB expression in MCF-7 with or without 10 nM E 2 in estrogen-deprived MCF-7 cells for 72 h, or in short-term oestrogen deprivation (STED) MCF-7 cells for 7 days and parental cells. (E) The schematic of HNRNPAB promoter region sequence selected from the transcription start site of -2 kb~+100 bp. (F) MCF-7 cells were transfected with pGL3 basic reporter vector containing HNRNPAB promoter, and pRL-TK reporter control vector containing luciferase activity, as well as control siRNA or different siRNA sequences of JUN , FOS , FOXA1 and SP1 . The relative fluorescence activity was measured. (G) CPTAC database analyzed the relative HNRNPAB and SP1 protein level in 68 ER+ BC patients. P value was determined by Pearson correlation analysis. (H) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector containing different regions of HNRNPAB promoter. The relative fluorescence activity was measured. (I) The schematic of three predicted conserved SP1 binding sites on the upstream -425 bp~+100 bp regions of the human HNRNPAB gene. Red boxes represent the predicted SP1 binding sites. (J) The DNA regions enriched by SP1 in ChIP assay were verified by PCR. (K) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector or containing “b” or “b” mutant regions. The relative fluorescence activity was measured. (L) The relative expression of SP1 and HNRNPAB mRNAs in MCF-7 cells bearing control or SP1 shRNAs analyzed by qRT-PCR. (M) Immunoblot assessment of SP1 and HNRNPAB expression in MCF-7 cells with or without SP1 shRNAs in the presence or absence of 10 nM E 2 for 48 h. Data was shown as mean ± S.D. from three independent experiments. Unpaired two-tailed Student's t test (C, H, K) and one-way ANOVA followed by Tukey's multiple comparisons test (F, L). ns, P >0.05; ***, P <0.001; ****, P <0.0001.

    Journal: International Journal of Biological Sciences

    Article Title: The Interaction of CircESR1 and HNRNPAB Regulates Cell Cycle Transition of Breast Cancer Cell

    doi: 10.7150/ijbs.126014

    Figure Lengend Snippet: Estrogen promotes HNRNPAB expression via SP1. (A) Immunoblot assessment of HNRNPAB in 2 human normal breast epithelial cell lines and 11 human BC cell lines. (B) CPTAC database analyzed the relative HNRNPAB and ERα protein level in 105 BC patients. P value was determined by Pearson correlation analysis. (C) The relative expression of TFF1 and HNRNPAB mRNAs after stimulated with 10 nM E 2 in estrogen-deprived MCF-7 cells for 48 h analyzed by qRT-PCR. (D) Immunoblot assessment of HNRNPAB expression in MCF-7 with or without 10 nM E 2 in estrogen-deprived MCF-7 cells for 72 h, or in short-term oestrogen deprivation (STED) MCF-7 cells for 7 days and parental cells. (E) The schematic of HNRNPAB promoter region sequence selected from the transcription start site of -2 kb~+100 bp. (F) MCF-7 cells were transfected with pGL3 basic reporter vector containing HNRNPAB promoter, and pRL-TK reporter control vector containing luciferase activity, as well as control siRNA or different siRNA sequences of JUN , FOS , FOXA1 and SP1 . The relative fluorescence activity was measured. (G) CPTAC database analyzed the relative HNRNPAB and SP1 protein level in 68 ER+ BC patients. P value was determined by Pearson correlation analysis. (H) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector containing different regions of HNRNPAB promoter. The relative fluorescence activity was measured. (I) The schematic of three predicted conserved SP1 binding sites on the upstream -425 bp~+100 bp regions of the human HNRNPAB gene. Red boxes represent the predicted SP1 binding sites. (J) The DNA regions enriched by SP1 in ChIP assay were verified by PCR. (K) 293T cells were transfected with control or SP1 vector, and pRL-TK reporter control vector containing luciferase activity, as well as pGL3 basic reporter vector or containing “b” or “b” mutant regions. The relative fluorescence activity was measured. (L) The relative expression of SP1 and HNRNPAB mRNAs in MCF-7 cells bearing control or SP1 shRNAs analyzed by qRT-PCR. (M) Immunoblot assessment of SP1 and HNRNPAB expression in MCF-7 cells with or without SP1 shRNAs in the presence or absence of 10 nM E 2 for 48 h. Data was shown as mean ± S.D. from three independent experiments. Unpaired two-tailed Student's t test (C, H, K) and one-way ANOVA followed by Tukey's multiple comparisons test (F, L). ns, P >0.05; ***, P <0.001; ****, P <0.0001.

    Article Snippet: For transcription factor mediated HNRNPAB expression, 0.2 μg pGL3 Basic luciferase reporter (RRID: Addgene_48743), 50 nmol JUN or FOS or FOXA1 or SP1 siRNAs and 0.02 μg pRL-TK plasmid (RRID: Addgene_11313) were transfected into cells using Lipofectamine 2000 (Invitrogen, Carlsbad, USA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Sequencing, Transfection, Plasmid Preparation, Control, Luciferase, Activity Assay, Fluorescence, Binding Assay, Mutagenesis, Two Tailed Test