Journal: Immunity & Ageing : I & A

Article Title: Cell autonomous expression of inflammatory genes in biologically aged fibroblasts associated with elevated NF-kappaB activity

doi: 10.1186/1742-4933-5-5

Figure Lengend Snippet: NF-κB activity. Shown are the results of a DNA binding assay of the NF-κB p65 transcription factor, using two biological replicates and four readouts for each sample. NF-κB activity, a key mediator of inflammation, is elevated in the nuclear fractions of fibroblasts from the group of older donors compared to young donors (p < 0.005). The DNA binding activity corresponds to a "low-grade" inflammatory state compared to the TNFα treated positive control.

Article Snippet: 10 μg of the nuclear cell fractions were used from each cell-line and their NF-κB p65 DNA binding activity was determined using the EZ-Detect NF-κB p65 Transcription Factor Kit (Pierce Biotechnology, Product No. 89859, Rockford, IL) according to the manufacturer's protocol.

Techniques: Activity Assay, DNA Binding Assay, Binding Assay, Positive Control

Journal: Immunity & Ageing : I & A

Article Title: Cell autonomous expression of inflammatory genes in biologically aged fibroblasts associated with elevated NF-kappaB activity

doi: 10.1186/1742-4933-5-5

Figure Lengend Snippet: List of inflammatory genes. Given are accession numbers of genes, fold changes and p-values between the group of older and young donors, a Pearson correlation coefficient (R) expressing similarity to the NF-κB profile as seen in Figure 1, and a description of genes.

Article Snippet: 10 μg of the nuclear cell fractions were used from each cell-line and their NF-κB p65 DNA binding activity was determined using the EZ-Detect NF-κB p65 Transcription Factor Kit (Pierce Biotechnology, Product No. 89859, Rockford, IL) according to the manufacturer's protocol.

Techniques: Expressing, Variant Assay, Binding Assay

Journal: Cells

Article Title: Protective Effects of SIRT6 Overexpression against DSS-Induced Colitis in Mice

doi: 10.3390/cells9061513

Figure Lengend Snippet: SIRT6 regulated expression and phosphorylation of TAK1. ( a ) The expression and phosphorylation levels of TAK1 were determined by immunohistochemistry. ( b , c ) The statistical results of ( a ). n = 3–6, ** p < 0.01.

Article Snippet: Sections then were blocked in 5% rat serum and incubated overnight at 4 °C with the diluted biotinylated primary antibody (1:500; rat-anti-mouse; anti-TAK1, #ab109526, Abcam, Cambridge, England, UK; anti-p-TAK1, #4508, Cell Signaling Technology, Danvers, MA, USA; anti-c-Jun, #ab40766, Abcam; anti-p-Jun, #ab32385, Abcam; anti-NF-κB p65 antibody, #ab32536, Abcam; anti-NF-κB p65 (phosphor S536), #ab86299, Abcam; anti-NF-κB p65 (acetyl K310) antibody, #ab19870, Abcam).

Techniques: Expressing, Immunohistochemistry

Journal: Life sciences

Article Title: Antioxidant and immunomodulatory activity induced by stevioside in liver damage: in vivo, in vitro and in silico assays.

doi: 10.1016/j.lfs.2019.03.035

Figure Lengend Snippet: Representative p65 immunohistochemistry obtained from livers of control rats (A), TAA-treated rats (B), STV+TAA-treated rats (C), and rats administered STV alone (D). The histogram depicts the percentage of p65 positivity obtained in immunohistochemistry sections (E). Increases in relative mRNA expression and protein levels of p65 (F, G) are shown. β-Actin was used as a control. Each bar represents the mean value of experiments conducted in duplicate assays ± SEM (n = 3). ap < 0.05 vs control. bp < 0.05 vs TAA.

Article Snippet: Taq polymerase (TaqMan, Universal Master Mix Prod # 4304437, Applied Biosystems, CA, USA) was used for the analysis of the relative expression of NF-κB (Rn01502266_m1), IL-1β (Rn00580432_m1), TNF-α (Rn01525859_g1), IL-6 (Rn01410330_m1), and IL-10 (Rn00563409_m1).

Techniques: Immunohistochemistry, Control, Expressing

Journal: Life sciences

Article Title: Antioxidant and immunomodulatory activity induced by stevioside in liver damage: in vivo, in vitro and in silico assays.

doi: 10.1016/j.lfs.2019.03.035

Figure Lengend Snippet: Genes studied and probes used in the in vitro qRT-PCR experiments.

Article Snippet: Taq polymerase (TaqMan, Universal Master Mix Prod # 4304437, Applied Biosystems, CA, USA) was used for the analysis of the relative expression of NF-κB (Rn01502266_m1), IL-1β (Rn00580432_m1), TNF-α (Rn01525859_g1), IL-6 (Rn01410330_m1), and IL-10 (Rn00563409_m1).

Techniques: In Vitro, Sequencing