Journal: Journal of Clinical Investigation

Article Title: TET repression and increased DNMT activity synergistically induce aberrant DNA methylation

doi: 10.1172/jci124070

Figure Lengend Snippet: Figure 3. Increased RELA binding levels at pro- moter regions of TET-targeting miRNAs. (A) Acti- vation of NF-κB signaling pathway in NUGC-3 cells by TNF-α. A downstream target gene of NF-κB signaling pathway, IL6, was upregulated by TNF-α treatment. Data represent mean ± SE. (B) Heat- map of RELA binding levels in NUGC-3 cells treated with TNF-α. RELA binding levels at genomic regions around TSSs of 44,112 transcripts were aligned according to the binding level after TNF-α treatment. Clear increase by the treatment was observed. Each row shows ± 2.5 kb centered on TSS. (C) Enriched motifs in RELA peaks detected in NUGC-3 cells treated with TNF-α. NF-κB binding motifs were most significantly enriched in NUGC-3 cells treated with TNF-α, showing successful detection of RELA binding sites. (D–F) RELA bind- ing status around the putative promoter regions of TET-targeting miRNAs. RELA binding levels at putative promoter regions of MIR26B (CTDSP1) (D) and MIR20A (MIR17HG) (E) were robustly increased by TNF-α treatment. RELA binding levels at these host genes were comparable to that at the IL6 promoter (F). Black boxes indicate genomic regions with peaks detected. The y axis represents the read pileup normalized to the total number of reads at a base pair position (rpm/bp).

Article Snippet: Crosslinked chromatin (100 μg) extracted from NUGC-3 cells with mock and TNF-α (30 ng/mL) treatment was immunoprecipitated using 5 μg antibody against RELA (R&D Systems, catalog AF5078).

Techniques: Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD80/CD86 costimulation regulates acute vascular rejection.

doi: 10.4049/jimmunol.175.9.6197

Figure Lengend Snippet: FIGURE 1. Onset of Ab-driven AVR at POD 5 in the absence of CD80 costimulation and onset of CMR at POD 17 in the absence of CD86 costimulation. A, Pa- thology of heart xenografts. HPS staining of xenograft hearts in CD80/ (POD 5), CD86/ (POD 6 and 17), and WT (POD 6 and 21) recipients. Scale bar, 100 m. Arrows indicate thrombosis and arrowheads hemor- rhage. B–D, Quantitation of serum xenoreactive Abs. Serum mouse anti-rat IgM (B), IgG1 (C), and IgG2a (D) Abs were measured in CD80/ (POD 0 and 5), CD86/ (POD 0, 6, and 17), and WT (POD 0, 6, and 21) recipients by flow cytometry. Data represent mean fluorescent intensity SD (n 4). , p 0.05 vs CD86/ POD 6, , p 0.01 vs CD86/ POD 6. POD 5 is also the endpoint of rejection in CD80/

Article Snippet: Sections were stained using goat anti-mouse IgM or IgG-biotin (Caltag Laboratories), CD4-biotin (YTS 191.1.2; Cedarlane Laboratories), CD8-biotin (53-6.7; BD Biosciences), or biotin-rat IgG2a or IgG2b (Cedarlane Laboratories).

Techniques: Staining, Quantitation Assay, Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD80/CD86 costimulation regulates acute vascular rejection.

doi: 10.4049/jimmunol.175.9.6197

Figure Lengend Snippet: FIGURE 2. Intragraft IgM and IgG deposition. Enumeration of intragraft IgM (A), and IgG (B) deposition in CD80/ (POD 5), CD86/ (POD 6 and 17), and WT (POD 6 and 21) grafts as measured by immunohisto- chemical staining (n 6). IgM and IgG deposition shown as number of positive cells/20 field SD. , p 0.01 vs CD86/ POD 6. ‡, POD 5 is also the endpoint of rejection in CD80/ mice.

Article Snippet: Sections were stained using goat anti-mouse IgM or IgG-biotin (Caltag Laboratories), CD4-biotin (YTS 191.1.2; Cedarlane Laboratories), CD8-biotin (53-6.7; BD Biosciences), or biotin-rat IgG2a or IgG2b (Cedarlane Laboratories).

Techniques: Staining