Journal: Frontiers in Microbiology

Article Title: NF-κBp50 and HDAC1 Interaction Is Implicated in the Host Tolerance to Infection Mediated by the Bacterial Quorum Sensing Signal 2-Aminoacetophenone

doi: 10.3389/fmicb.2017.01211

Figure Lengend Snippet: Pharmacological and genetic inhibition of HDAC1 rescue the 2-AA-mediated immunomodulation. (A) Schematic representation of the 2-AA treatments. The human monocytes THP-1 cells and mouse macrophage RAW264.7 cell cells were left untreated (No Pre: No Tolerization) or pretreated with 2-AA for 24 h or 48 h (2-AA Pre: 2-AA tolerization) respectively, and then stimulated (Sti) with 2-AA. (B) Expression of TNF (1 h) was measured in non-pretreated, 2-AA pretreated and 2-AA + TSA pretreated THP-1 cells following 2-AA stimulation. Transcript levels were assessed by qRT-PCR and normalized to GAPDH . (C) ChIP assay of H3K18ac at the TNF promoter of 2-AA-pretreated, 2-AA and TSA pretreated or non-pretreated THP-1 cells following 3 h 2-AA stimulation, assessed by qRT-PCR with primer covering the promoter site region of TNF relative to GAPDH. (D,E) ELISA of IL-1β and Mcp1 secretion in culture supernatant of 2-AA pretreated vector control RAW264.7 and HDAC1 KD cells following 6 h 2-AA stimulation ( n = 3; means ± SDs; p < 0.05, Student’s t -test).

Article Snippet: The binding of a 2-AA treated nuclear fraction was used to identify the substrates of histone modifying enzymes using Pre-Sure Histone H3 peptide Array ELISA kit (EpiGentek) according to the manufactures protocol.

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Control

Journal: Frontiers in Microbiology

Article Title: NF-κBp50 and HDAC1 Interaction Is Implicated in the Host Tolerance to Infection Mediated by the Bacterial Quorum Sensing Signal 2-Aminoacetophenone

doi: 10.3389/fmicb.2017.01211

Figure Lengend Snippet: 2-AA modulates H3K18 acetylation. Screening for Histone H3 acetylation following 2-AA stimulation. Nuclear extraction was prepared from THP-1 cells 1 h post-stimulation with 2-AA. The interaction of the enzyme in the sample solution with the acetylated H3K peptides was measured by using Histone H3 Peptide Array ELISA kit (number of replicates = 2; means ± SDs).

Article Snippet: The binding of a 2-AA treated nuclear fraction was used to identify the substrates of histone modifying enzymes using Pre-Sure Histone H3 peptide Array ELISA kit (EpiGentek) according to the manufactures protocol.

Techniques: Extraction, Peptide Microarray, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Haplotyping the human leukocyte antigen system from single chromosomes

doi: 10.1038/srep30381

Figure Lengend Snippet: ( a ) Bivariate flow plot of single chromosome acquisition sorting. The X-axis indicates the fluorescence emitted by Chromomycin A3 specific binding to guanine-cytosine (C-G) regions of DNA, the Y-axis indicates the fluorescence emitted by Hoechst 33258 binding to adenine-thymine (A-T) regions of DNA. ( b ) 24 Sure (+) BAC microarray (BlueGnome, Cambridge) of chromosome 6 signal hybridised against male reference genomic DNA indicating the presence of chromosomal DNA from chromosome 6. Background noise for the remaining chromosomes indicates a lack of DNA from chromosomes 1–5, 7–22, X or Y.

Article Snippet: Samples were then run on 24 Sure (+) BAC microarray (BlueGnome, Cambridge) to select samples containing DNA from flow-sorted then amplified chromosome 6 samples.

Techniques: Fluorescence, Binding Assay, Microarray