Journal: PLoS ONE

Article Title: ZFP36L1 Negatively Regulates Plasmacytoid Differentiation of BCL1 Cells by Targeting BLIMP1 mRNA

doi: 10.1371/journal.pone.0052187

Figure Lengend Snippet: (A) Heat map expression profile of known ZFP36L1 target genes in terminal B cell differentiation. The profiles are compared with the reference genes, ZFP36L1, BLIMP1 and XBP1 (first three rows). GMCSF, VEGFA and IL-3 are validated ZFP36L1 targets; the remaining genes have been validated for ZFP36 . Data was taken from GEO Accession number GSE 6691 . Red indicates high and blue, low expression. Key: CLL: B chronic lymphocytic leukemia, MM: multiple myeloma, WM-BL: Waldenstrom’s macroglobulinemia B cells, WM-PB: Waldenstrom’s macroglobulinemia plasma cells, NBC: normal B cells, PC: normal plasma cells. (B) Graphical representation of the ARACNe network for BLIMP1 and ZFP36L1. Nodes representing the BLIMP1 and ZFP36L1 hubs are shown enlarged. Only first neighbours of these hubs are shown in the module. Nodes representing inferred BLIMP1 targets (significantly up-regulated in normal B cells) are blue; nodes representing inferred ZFP36L1 targets (significantly up-regulated in normal plasma cells) are red. Network graphics were generated as group attribute layout using Cytoscape version 2.6.0. .

Article Snippet: Nitrocellulose membranes were incubated with appropriate dilution of the following primary antibodies for 1h; rabbit polyclonal anti-BRF1/2 (ZFP36L1/2) (Cell Signalling, Danvers, MA, USA), goat anti-ZFP36 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-BLIMP1 (Cell Signalling).

Techniques: Expressing, Cell Differentiation, Generated

Journal: The Journal of Clinical Investigation

Article Title: RNA-binding protein ZFP36L1 maintains posttranscriptional regulation of bile acid metabolism

doi: 10.1172/JCI94029

Figure Lengend Snippet: (A) FXR ChIP-seq analysis (21) of the Zfp36l1 genomic loci showing putative FXREs. (B) IHHs were transfected with a plasmid containing the Zfp36l1 promoter region (2 kb) upstream of a luciferase reporter in combination with increasing amounts of a mouse Fxrα2 expression plasmid. Cells were treated with vehicle or GW4064 (GW) (1 μM) for 24 hours after transfection. Luciferase activity was normalized to β-gal and is expressed as the fold change (n = 6 wells/condition). ZFP36L1 mRNA (C) and protein (D and E) in C57BL/6 WT and Fxr−/− mice treated daily with vehicle, GW4064, or GSK2324 (60 mpk) for 3 days (n = 7–9 mice/group). (F) ZFP36L1 mRNA and protein in C57BL/6 WT mice fed either a control (Ctr) or 0.5% CA diet. Gene expression analysis was determined by qRT-PCR and normalized to Tbp, and protein analysis was determined by Western blotting, with PDI used as a loading control. Data represent the mean ± SEM. **P < 0.01 and ***P < 0.001, by 1-way ANOVA (B and C) and Student’s t test (F). Veh, vehicle.

Article Snippet: Anti-ZFP36L1 antibody (BRF1/2, catalog 2119; Cell Signaling Technology; 1:1,000); anti-CYP7A1 antibody (catalog MABD42; Sigma-Aldrich; 1:1,000); protein disulfide isomerase (PDI) (catalog 3501; Cell Signaling Technology; 1:1,000); and secondary anti-rabbit HRP-conjugated antibody (GE Healthcare; 1:10,000) were used.

Techniques: ChIP-sequencing, Transfection, Plasmid Preparation, Luciferase, Expressing, Activity Assay, Quantitative RT-PCR, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: RNA-binding protein ZFP36L1 maintains posttranscriptional regulation of bile acid metabolism

doi: 10.1172/JCI94029

Figure Lengend Snippet: (A, C, and E) Zfp36l1 mRNA and (B, D, and F) ZFP36L1 protein in the livers of (A and B) WT and Fxr−/− mice (n = 16–8 WT; n = 7–6 Fxr–/– mice/group); (C and D) littermate Fxrfl/fl and FxrL-KO mice (n = 13–7 WT; n = 11–7 FxrL-KO mice/group); and (E and F) Shp−/− mice (n = 8–7 mice/group) treated with either vehicle or GSK2324 for 30 minutes, 1 hour, or 2 hours. Gene expression analysis was determined by qRT-PCR and normalized to Tbp, and protein analysis was determined by Western blotting, with PDI used as a loading control. Data represent the mean ± SEM. **P < 0.01 and ***P < 0.001, by 1-way ANOVA. MW, molecular weight.

Article Snippet: Anti-ZFP36L1 antibody (BRF1/2, catalog 2119; Cell Signaling Technology; 1:1,000); anti-CYP7A1 antibody (catalog MABD42; Sigma-Aldrich; 1:1,000); protein disulfide isomerase (PDI) (catalog 3501; Cell Signaling Technology; 1:1,000); and secondary anti-rabbit HRP-conjugated antibody (GE Healthcare; 1:10,000) were used.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Molecular Weight

Journal: The Journal of Clinical Investigation

Article Title: RNA-binding protein ZFP36L1 maintains posttranscriptional regulation of bile acid metabolism

doi: 10.1172/JCI94029

Figure Lengend Snippet: (A) Mouse and (B) human (WT and mutant) Cyp7a1 3′-UTR cloned downstream of a luciferase reporter and then transfected in IHHs in combination with increasing amounts of a mouse Zfp36l1 expression plasmid. Luciferase activity was determined after 24 hours and normalized to β-gal and is expressed as the fold change (n = 6 wells/condition). (C) ZFP36L1 mRNA and protein, (D) CYP7A1 mRNA and protein, and (E) mRNA levels of bile acid synthesis genes in male C57BL/6 WT mice treated with either adenovirus-control (Ad-Ctr) or Ad-Zfp36l1 (n = 10 mice/group). (F) Bile acid concentration in gall bladders and (G) plasma cholesterol levels in Ad-Ctr– or Ad-Zfp36l1–treated mice (n = 10 mice/group). Gene expression analysis was determined by qRT-PCR and normalized to Tbp. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, by 1-way ANOVA (A and B) and Student’s t test ( C–G).

Article Snippet: Anti-ZFP36L1 antibody (BRF1/2, catalog 2119; Cell Signaling Technology; 1:1,000); anti-CYP7A1 antibody (catalog MABD42; Sigma-Aldrich; 1:1,000); protein disulfide isomerase (PDI) (catalog 3501; Cell Signaling Technology; 1:1,000); and secondary anti-rabbit HRP-conjugated antibody (GE Healthcare; 1:10,000) were used.

Techniques: Mutagenesis, Clone Assay, Luciferase, Transfection, Expressing, Plasmid Preparation, Activity Assay, Concentration Assay, Quantitative RT-PCR

Journal: The Journal of Clinical Investigation

Article Title: RNA-binding protein ZFP36L1 maintains posttranscriptional regulation of bile acid metabolism

doi: 10.1172/JCI94029

Figure Lengend Snippet: (A) ZFP36L1 mRNA and protein, (B) CYP7A1 mRNA and protein, and (C) mRNA levels of bile acid synthesis genes in male littermate Zfp36l1fl/fl (n = 9) and Zfp36l1L-KO mice (n = 11). (D) Biliary bile acid concentration and bile acid composition and (E) plasma total and HDL cholesterol levels in littermate Zfp36l1fl/fl and Zfp36l1L-KO mice. Gene expression analysis was determined by qRT-PCR and normalized to Tbp. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, by Student’s t test.

Article Snippet: Anti-ZFP36L1 antibody (BRF1/2, catalog 2119; Cell Signaling Technology; 1:1,000); anti-CYP7A1 antibody (catalog MABD42; Sigma-Aldrich; 1:1,000); protein disulfide isomerase (PDI) (catalog 3501; Cell Signaling Technology; 1:1,000); and secondary anti-rabbit HRP-conjugated antibody (GE Healthcare; 1:10,000) were used.

Techniques: Concentration Assay, Expressing, Quantitative RT-PCR

Journal: The Journal of Clinical Investigation

Article Title: RNA-binding protein ZFP36L1 maintains posttranscriptional regulation of bile acid metabolism

doi: 10.1172/JCI94029

Figure Lengend Snippet: (A–C) Hepatic expression of Shp, Zfp36l1, and Cyp7a1 genes in Zfp36l1fl/fl and littermate Zfp36l1L-KO mice treated with either vehicle (n = 10, Zfp36l1fl/fl; n = 13, Zfp36l1L-KO) or GSK2324 for 30 minutes (n = 8, Zfp36l1fl/fl; n = 7, Zfp36l1L-KO), 1 hour (n = 8, Zfp36l1fl/fl; n = 8, Zfp36l1L-KO), or 2 hours (n = 8, Zfp36l1fl/fl; n = 7, Zfp36l1L-KO) before sacrifice. Gene expression analysis was determined by qRT-PCR and normalized to Tbp. (D) Cyp7a1 levels after GSK treatment in Zfp36l1fl/fl and Zfp36l1L-KO mice compared with levels in vehicle-treated mice, set to 1 (blue lines represent the same data as in C). (E) Summary diagram depicting known FXR-regulated pathways that control bile acid synthesis feedback inhibition. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, by 1-way ANOVA.

Article Snippet: Anti-ZFP36L1 antibody (BRF1/2, catalog 2119; Cell Signaling Technology; 1:1,000); anti-CYP7A1 antibody (catalog MABD42; Sigma-Aldrich; 1:1,000); protein disulfide isomerase (PDI) (catalog 3501; Cell Signaling Technology; 1:1,000); and secondary anti-rabbit HRP-conjugated antibody (GE Healthcare; 1:10,000) were used.

Techniques: Expressing, Quantitative RT-PCR, Inhibition