Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: TTP overexpression represses ERα, PR, GR and AR transactivation in MCF-7 cells. Human breast cancer MCF-7 cells were transiently transfected with empty pCMV-3TAG vector (control) or with 0.25 μg of pCMV-3TAG-TTP (FLAG-TTP) along with the corresponding luciferase reporter vector. The effect of TTP on nuclear hormone receptors transactivation was determined by assay of luciferase activity, as described under “Experimental Procedures.” Assays were performed in triplicate in three independent experiments in the presence (white bars) or absence (black bars) of the corresponding nuclear receptor ligands (17β-estradiol for ERα, progesterone for PR, 5α-Androstan-17β-ol-3-on for AR and dexamethasone for GR. The results are represented as mean ± S.E. Differences in ERα activity between control MCF-7 cells and TTP-expressing MCF-7 cells are statistically significant ( p < 0.05).

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Luciferase, Activity Assay, Expressing

Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: TTP interacts with ERα, PR, GR and AR in vivo . Endogenous ERα (A), PR (B), AR (C) and GR (C) were immunoprecipitated from protein extracts prepared from MCF-7 cells using specific antibodies. Immunoprecipitated proteins were resolved by PAGE, and the binding of TTP was visualized by Western blot. The input lane represents 10% of the protein extract used in the coimmunoprecipitation assays. IP: immunoprecipitation, WB: Western blot.

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: In Vivo, Immunoprecipitation, Binding Assay, Western Blot

Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: siRNA-mediated knockdown of TTP protein levels in MCF-7 cells. MCF-7 cells were transfected with specific siRNA-TTP (1.25 μg) or with an unrelated control siRNA (2.5 μg). (A) Protein extracts from MCF-7 cultures were resolved by PAGE, and expression levels of TTP, ERα, PR, GR, AR and tubulin, as a loading control protein, were evaluated by Western blot using specific antibodies as described under “Experimental Procedures.” TTP knockdown increases ERα, PR, GR and AR transactivation. MCF-7 cells were transfected with an unrelated siRNA (control) or a specific TTP-siRNA (1.5 μg), along with TK-LUC, in the presence of the corresponding nuclear receptor ligands. The effect of TTP-siRNA on ERα (B), PR (C), GR (D) and AR (E) transactivation was determined by a luciferase assay and compared with luciferase activity in MCF-7 cells transfected with empty pCMV-3TAG vector and the corresponding Tk-LUC reporter vector. Results, in triplicate in three independent experiments, are represented as mean ± S.E. (error bars). Differences in ERα activity in MCF-7 cells transfected with TTP or with TTP-siRNA were shown to be statistically significant ( p < 0.05).

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: Knockdown, Transfection, Control, Expressing, Western Blot, Luciferase, Activity Assay, Plasmid Preparation

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: The highly expressed TTP was identified as a crucial factor in response to the IgE-mediated inflammatory syndrome. A , the robust rank aggregation (RRA) algorithm integrated upregulated genes (Log 2 Foldchange>1 and p < 0.05) from three datasets of three IgE-mediated inflammatory syndromes (allergic rhinitis, asthma, and atopic dermatitis) and their corresponding normal samples. B , the ranking graph displayed the genes that were upregulated in at least two datasets. Among them, the genes corresponding to the red dots (Frequency = 3) showed a significant increase in all three disease datasets, including TTP. C , intersection of the genes encoding CCCH-ZF (zinc finger)-type proteins with the genes upregulated in the nasal mucosa of patients with allergic rhinitis. D , the assessment of evolutionary conservation of amino acid positions, especially two CCCH-ZF repeat regions, in the mouse TTP protein. Conservation scores ranged from 1 to 9, where one was the most highly variable, five was of intermediate conservation, and nine was the most highly conserved position. E , experimental scheme of ragweed pollen (RW)-induced AR model, divided into the sensitization stage (0 and 7 days) and the stimulation stage (14–17 days). The control mice were treated with PBS instead of RW (N = 6 per group). F , relative RNA expressions of TTP in nasal tissues of RW-induced AR mice and control mice. G , compare the frequencies of sneezing and nose-rubbing within 10 min between control mice and AR mice after 14 days. H , immunofluorescence staining of TTP protein ( red ) and CD4 protein ( green ) was performed in normal and RW nasal tissues. DAPI ( blue ) was used for nuclear staining (Scale bar = 50 μm). I , quantitative analyses of CD4 + T cell percentages and TTP fluorescence intensity in nasal mucosal tissues from sham and AR model mice. Data are shown as means ± SD (N = 6), Student's t test was used for statistical analysis of control mice and AR mice.

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Control, Immunofluorescence, Staining, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: TTP suppressed T helper 2 (Th2)-type inflammation in AR mice. A , schematics of splenocytes isolation from AR-mice and the differentiation of splenic naive CD4+ T cells model in vitro . B , the levels of Interleukin (IL)-4, IL-5, and IL-13, which belong to the Th2 cytokine family, were measured by ELISA assays in the cell supernatant. The cell supernatant was collected from mononuclear cells of the spleen that had been treated with ionomycin and phorbol 12-myristate 13-acetate (PMA) for 6 h. C , after treatment with ionomycin, PMA, and GolgiPlug, the cells with the double stain of anti-CD4 and anti-IL-4-APC were detected by flow cytometry to analyze the percentage of Th2 cells. Data are shown as means ± SD (N = 3 or 6). One-way analysis of variance (ANOVA) was used for statistical analysis of control mice, AR mice, and AR mice transduced with control vectors or TTP overexpression vectors.

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Isolation, In Vitro, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry, Control, Transduction, Over Expression

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: TTP overexpression inhibited Th2 differentiation in vitro . A , schematics of naive CD4+ isolation, activation, and Th2 differentiation. B , a representative flow cytometric plot and the percentage of Th2 cells was detected using flow cytometry. C , the protein and mRNA expression of TTP in CD4+ T cells. D , the protein and mRNA expression of TTP in Th2 cells after virus infection. E , the levels of IL-5 were detected using real-time PCR and ELISA, respectively, in Th2 cells after virus infection. F , the percentage of Th2 cells was detected using flow cytometry in Th2 cells after virus infection. Data are shown as means ± SD (N = 3). Student's t test and one-way ANOVA were used for statistical analysis.

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Over Expression, In Vitro, Isolation, Activation Assay, Flow Cytometry, Expressing, Virus, Infection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: TTP knockdown exacerbated allergic phenotypes and promoted Th2-type inflammation in AR mice. A , experimental scheme of RW-induced AR mice with or without TTP knockdown via intravenous injection of lentiviral vectors (Lv-shNC or Lv-shTTP). B , expression of TTP mRNA in AR mice with or without TTP knockdown. C , immunofluorescence staining for TTP protein ( red ) and CD4 protein ( green ) in normal and RW-exposed nasal tissues. DAPI ( blue ) was used for nuclear staining (Scale bar = 50 μm). D , quantitative analysis of TTP fluorescence intensity in nasal mucosal tissues from lentivirus-injected mice. E , hematoxylin-eosin (H&E)-stained nasal mucosal tissue sections, showing nasal mucosa thickness (Scale bar = 100 μm). Dashed lines indicate the boundary between the epithelial layer and lamina propria. F and G , changes in the number of sneezes and nasal rubbings after TTP knockdown in AR mice. H , schematic of splenocyte isolation from AR mice and in vitro treatment. I – K , Levels of IL-4, IL-5, and IL-13 in the cell culture supernatant of splenocytes treated with ionomycin and PMA for 6 h. Data are presented as means ± SD (N = 3 or 6). Student's t test was used for statistical analysis of AR mice transduced with shNC or shTTP.

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Knockdown, Injection, Expressing, Immunofluorescence, Staining, Fluorescence, Isolation, In Vitro, Cell Culture, Transduction

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: Transcriptional profiles of CD4+ T cell isolated from AR mouse spleen reveal categories of genes associated with TTP. A , RNA transcriptome microarray analysis of CD4+ T cell isolated from mouse. B , a volcano plot of all DEGs between CD4+ T cells isolated from LV-TTP and LV-Vector infected AR mouse. C , GSEA analysis identified significant enrichment of signaling pathways based on KEGG and GO analysis following TTP overexpression. D , KEGG pathway enrichment and GO functional classification analysis of DEGs. Data are shown as means ± SD (N = 3).

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Isolation, Microarray, Plasmid Preparation, Infection, Protein-Protein interactions, Over Expression, Functional Assay

Journal: The Journal of Biological Chemistry

Article Title: RNA-binding protein tristetraprolin inhibits Th2 cell activation and differentiation in allergic rhinitis by promoting TRIM18 mRNA decay

doi: 10.1016/j.jbc.2026.111240

Figure Lengend Snippet: The TTP protein acted as a negative regulator of TRIM18-mediated Th2-type inflammation by reducing TRIM18 mRNA stability. A , protein–protein interaction network of 16 potential substrates of E3 ubiquitin ligase TRIM18, constructed using the STRING database and supplemented with functional annotations via GO (Gene Ontology) pathway enrichment analyses, highlighting key biological processes and signaling pathways associated with these substrates. B and C , the qRT-PCR and ELISA were performed to detect the mRNA level and secreted protein concentration of IL-5, respectively, in naive CD4+ T cells or in vitro differentiated Th2 cells transfected with TRIM18 overexpression vector or empty vector. D , schematic diagram of the experimental workflow: Naive CD4+ T cells were induced to differentiate into Th2 cells under in vitro and were co-transfected with combinations of TTP overexpression vector, TRIM18 overexpression vector, or corresponding empty vectors. E , the mRNA level of TRIM18 in differentiated Th2 cells from the three groups (Control, TTP-OE, TTP+TRIM18-OE) was detected by qRT-PCR assays. F and G , The mRNA level and secreted protein concentration of IL-5 were measured by qRT-PCR and ELISA, respectively, in differentiated Th2 cells from the three groups (Control, TTP-OE, TTP+TRIM18-OE). H , flow cytometric analysis of the percentage of CD4+IL-4+ Th2 cells among the three groups after in vitro differentiation, and the proportion of double-positive cells was quantified to assess the impact of TTP and/or TRIM18 overexpression on Th2 cell differentiation. Data are shown as means ± SD (N = 3), One-way ANOVA was used for statistical analysis.

Article Snippet: Then, the sections were co-incubated with antibodies against TTP (1:100 dilution, Proteintech, Cat No.66938-1-Ig) and CD4 (1:50 dilution, Abclonal, Cat No.A0363) overnight at 4 °C.

Techniques: Ubiquitin Proteomics, Construct, Functional Assay, Protein-Protein interactions, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Protein Concentration, In Vitro, Transfection, Over Expression, Plasmid Preparation, Control, Cell Differentiation

Journal: Scientific Reports

Article Title: The Fragment HMGA2-sh-3p20 from HMGA2 mRNA 3′UTR Promotes the Growth of Hepatoma Cells by Upregulating HMGA2

doi: 10.1038/s41598-017-02311-0

Figure Lengend Snippet: HMGA2-sh-3p20 upregulates HMGA2 by blocking the TTP-mediated degradation of HMGA2 mRNA. ( A ) The relative expression of HMGA2-sh-3p20 was assessed by qRT-PCR in 35 pairs of clinical HCC tissues and corresponding peritumor tissues ( ***P < 0.001; Wilcoxon’s signed-rank test). ( B ) The correlation between HMGA2 mRNA levels and HMGA2-sh-3p20 levels was measured by qRT-PCR in 30 cases of clinical HCC tissues ( **P < 0.01, r = 0.586; Pearson’s correlation coefficient). ( C ) The luciferase activities of pGL3-HMGA2 were examined by luciferase reporter gene assays in HepG2 cells. ( D ) The expression of HMGA2 was assessed by qRT-PCR and Western blot analysis in Huh7 cells. ( E ) The diagram of TTP-mediated mRNA degradation. ( F ) The diagram of HMGA2-sh-3p20 antagonizes the interaction of TTP with non-hairpin within 3′UTR of HMGA2 mRNA. ( G ) TTP RIP-PCR of HMGA2 in HepG2 cells. ( H ) Effect of TTP on the expression of HMGA2 was measured by qRT-PCR and Western blot analysis in HepG2 cells. ( I ) Effect of HMGA2-sh-3p20 on the expression of TTP-mediated HMGA2 was assessed by qRT-PCR and Western blot analysis in HepG2 cells. The full length blots images are given as Supplementary Fig. . ( J ) TTP RIP-qPCR of HMGA2 in HepG2 cells transfected with HMGA2-sh-3p20. ( K ) Effect of HMGA2-sh-3p20 on the levels of HMGA2 mRNA in HepG2 cells transfected with si-TTP by qRT-PCR. ( L , M ) Effect of HMGA2-sh-3p20 on the half-life of HMGA2 mRNA in HepG2 cells ( L ) or HepG2 cells transfected with si-TTP ( M ) by qRT-PCR. Every experiment was repeated three times. Error bars represent s.d. (n = 3), **p < 0.01; ***p < 0.001 and not significant (NS), Student’s t test.

Article Snippet: The dilution of primary antibody is following: HMGA2 (1: 5000, Genetex), PTEN (1:800, Proteintech), TTP (1:1000, Proteintech), Drosha (1:1000, Proteintech), DGCR8 (1:800, Proteintech), Dicer (1:800, Proteintech), β-actin (1:2000, Abcam).

Techniques: Blocking Assay, Expressing, Quantitative RT-PCR, Luciferase, Western Blot, Transfection

Journal: Scientific Reports

Article Title: The Fragment HMGA2-sh-3p20 from HMGA2 mRNA 3′UTR Promotes the Growth of Hepatoma Cells by Upregulating HMGA2

doi: 10.1038/s41598-017-02311-0

Figure Lengend Snippet: A model shows that the fragment HMGA2-sh-3p20 from HMGA2 mRNA 3′UTR promotes the growth of hepatoma cells by upregulating HMGA2. Bioinformatics analysis shows that 3′UTR of HMGA2 mRNA contains the hairpin structure (HMGA2-sh). Drosha and DGCR8 cleave the HMGA2-sh from the 3′UTR of HMGA2 mRNA, and Dicer contributes to the generation of the HMGA2-sh-3p20 from the HMGA2-sh. Furthermore, HMGA2-sh-3p20 is able to increase the levels of HMGA2 by antagonizing TTP-mediated HMGA2 degradation, while it decreases PTEN by targeting 3′UTR of PTEN mRNA. In addition, the downregulated-PTEN is not able to depress the expression of HMGA2, leading to the upregulation of HMGA2. Functionally, HMGA2-sh-3p20-enhanced HMGA2 accelerates the growth of liver cancer cells.

Article Snippet: The dilution of primary antibody is following: HMGA2 (1: 5000, Genetex), PTEN (1:800, Proteintech), TTP (1:1000, Proteintech), Drosha (1:1000, Proteintech), DGCR8 (1:800, Proteintech), Dicer (1:800, Proteintech), β-actin (1:2000, Abcam).

Techniques: Expressing

Journal: Molecular cell

Article Title: Tandem deubiquitination and acetylation of SPRTN promotes DNA-protein crosslinks repair and protects against aging

doi: 10.1016/j.molcel.2020.06.027

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-VCPIP1 (C2C3) , Genetex , Cat# GTX107169; RRID:AB_1952546.

Techniques: Recombinant, Staining, Mutagenesis, In Situ, Fractionation, Software

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Expression of human α-tocopherol transfer protein (α-TTP) eukaryotic expression vector in Human Hepatoma cells (HepG2). ( A ) Result of quantitative real-time PCR; ( B ) Result of Western blot; ( C ) Result of Immunofluorescence (10×). pcDNA: pcDNA3.1-mycHisa empty vector; p-TTP: human α-TTP eukaryotic expression vector.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Expression levels of α-TTP related genes. SEC23A : Sec23 homolog A; RTP4 : receptor transporter protein 4; CLIC3 : chloride intracellular channel 3; CENPE : centromere protein E; KCNQ1 : potassium voltage-gated channel subfamily Q member 1; GOLGA4 : golgi autoantigen, golgin subfamily a, 4; BNIP3 : BCL2/adenovirus E1B 19 kDa interacting protein 3; CENPF : centromere protein F. Data are presented as mean ± standard error; the asterisk indicated significant difference between control and treatment group.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Composition of the ligation reaction.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Ligation, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Details of primers used for quantitative real-time PCR.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Sequencing