Journal: PLoS ONE

Article Title: Crosstalks of the PTPIP51 interactome revealed in Her2 amplified breast cancer cells by the novel small molecule LDC3/Dynarrestin

doi: 10.1371/journal.pone.0216642

Figure Lengend Snippet: Antibody list.

Article Snippet: IκBα , Recombinant Human IκB alpha/NFKBIA protein 02 , Mouse monoclonal , MM02 , 1:100 , Sino Biological Inc., North Wales, PA, USA Cat.# 12045-H07E.

Techniques: Recombinant, Purification

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for SRC-1/NF-kB association. The signals for p65, p50 and Pol II indicate that NF-kB was activated and mRNA synthesis was initiated. Actin signal is a control of DNA input. (B) Cotransfection of SRC-1 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 and p50 expression vectors, which were used at 0.1 μg/point for each plasmid DNA. (C) Immunoblotting of IkBα protein in HEK293 cells transfected by SRC-1. SRC-1 expression is confirmed in the transfected cells. Actin protein is a control for protein loading. (D) Cotransfection of SRC-1 with NF-kB luciferase reporter. The same condition was used as described in the legend for panel 2 except the reporter DNA.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Cotransfection, Expressing, Plasmid Preparation, Western Blot, Transfection, Luciferase

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for SRC-2/NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. The signals for p65, p50 and Pol II is presented in Fig. 1A. (B) Nuclear exclusion of SRC-2. The cytoplasmic and nuclear proteins were extracted from HEK293 cells after TNF-treatment. SRC-2 protein was quantified in the extracts by immunoblotting. Actin and SP1 protein signals are used as controls for protein loading of the cytoplasmic and nuclear extracts, respectively. (C) Cotransfection of SRC-2 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 expression vectors at 0.1 μg/point. (D) Immunoblotting of IkBα protein in HEK293 cells transfected. SRC-2 protein level is confirmed in cells transfected for overexpression or knockdown.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Western Blot, Cotransfection, Expressing, Transfection, Over Expression

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for SRC-3/NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. The signals for p65 and p50 demonstrates activation of NF-kB by TNF-α. (B) Cotransfection of SRC-3 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 expression vector at 0.1 μg/point. The SRC-3 DNA (μg) is indicated. (C) Knockdown of SRC-3 by vector-based RNAi expression. The DNA (μg) of SRC-3 RNAi vector is indicated. (D) Immunoblotting of IkBα protein in HEK293 cells transfected. SRC-3 protein level is confirmed in cells transfected for knockdown.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Activation Assay, Cotransfection, Expressing, Plasmid Preparation, Western Blot, Transfection

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for HDAC1/NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. (B) Cotransfection of HDAC1 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 expression vector at 0.1 μg/point. The HDAC1 DNA (μg) is indicated. (C) Knockdown of HDAC1 by vector-based RNAi expression. The DNA (μg) of HDAC1 RNAi (1RNAi) vector is indicated. (D) Immunoblotting of IkBα protein in HEK293 cells transfected. HDAC1 protein level is confirmed in cells transfected for overexpression knockdown.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Cotransfection, Expressing, Plasmid Preparation, Western Blot, Transfection, Over Expression

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for HDAC2/NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. (B) Cotransfection of HDAC2 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 expression vector, which was used at 0.1 μg/point. The HDAC2 DNA (μg) is indicated. (C) Knockdown of HDAC2 by vector-based RNAi expression. The DNA (μg) of HDAC2 RNAi vector is indicated. 2RNAi is abbreviation of HDAC2 RNAi. (D) Immunoblotting of IkBα protein in HEK293 cells transfected. The change in HDAC2 protein is confirmed in cells transfected for overexpression knockdown.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Cotransfection, Expressing, Plasmid Preparation, Western Blot, Transfection, Over Expression

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for HDAC3/NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. (B) Knockdown of HDAC3 by vector-based RNAi expression. The DNA (μg) of HDAC3 RNAi vector is indicated. (C) Cotransfection of HDAC3 with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 expression vector at 0.1 μg/point. The HDAC3 DNA (μg) is indicated.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Plasmid Preparation, Expressing, Cotransfection

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) ChIP assay for SMRT and NCoR interaction with NF-kB association. This result was obtained in the same condition as for ChIP assay of SRC-1. (B) Cotransfection of SMRT or NCoR with IkBα reporter. The IkBα reporter was activated by cotransfection of p65 and p50 expression vectors, which was used at 0.1 μg/point for each vector. Plasmid DNA (μg) for SMRT or NCoR is indicated. (C) Knockdown of SMRT or NCoR by vector-based RNAi expression. Vector DNA (μg) of RNAi expressing plasmid is indicated.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Cotransfection, Expressing, Plasmid Preparation

Journal:

Article Title: COACTIVATORS AND COREPRESSORS OF NF-KB IN IkB ALPHA GENE PROMOTER

doi: 10.1074/jbc.M500754200

Figure Lengend Snippet: (A) The ChIP assay was conducted as stated in the method section. To help readers in understanding the complex switches among the coactivators and corepressors, and between the coactivator and corepressors, ChIP data presented in Figures 1 through ​through77 are combined together here. (B) IkBα mRNA expression was induced by TNF-treatment in 293 cells. IkBα mRNA was determined by Taqman RT-PCR as stated in methods. (C) Regulation of IkBα mRNA by the coactivators and corepressors.

Article Snippet: The real time RT-PCR reaction was conducted in triplicates using Taqman IkBα probe (Hs00153283_m1, Applied Biosynthesis).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction