Journal: PLoS ONE

Article Title: Quercetin Suppresses Cyclooxygenase-2 Expression and Angiogenesis through Inactivation of P300 Signaling

doi: 10.1371/journal.pone.0022934

Figure Lengend Snippet: ( A–C ), The MDA-MB-231 cells were transfected with FLAG-p300 for 24 h and then treated with quercetin for 24 h. Nuclear extracts were prepared and p50 was immunoprecipitated with a p50 antibody. Acetylated p50 (Actyl-p50) ( A ) was analyzed by Western blots using an acetyl-lysine antibody. The binding of p50 to a biotinylated COX-2 promoter probe ( B ) and chromatin structure ( C ) were detected by streptavidin-agarose pulldown and ChIP assay, respectively. ( D ), MDA-MB-231 cells were treated with roscovitine (20 µM) or quercetin (100 µM) for 24 h, then transfected with p300-expressing vector. After 24 h, the HAT activity was measured. The empty vector was used as a transfection control. The figures are representatives of three experiments. Each bar represents mean ±SD of three experiments. *, P <0.05, significant differences between treatment groups and control groups. NSP, non-specific probe; EV, empty vector.

Article Snippet: After extensive washing, p50 proteins were separated in a 4–20% SDS-PAGE system and acetylated p50 was detected with a monoclonal antibody against acetyl-lysine (1∶1000 dilution) (Cell Signaling Tech., Beverly, MA).

Techniques: Transfection, Immunoprecipitation, Western Blot, Binding Assay, Expressing, Plasmid Preparation, Activity Assay

Journal: BMC Medicine

Article Title: Serum transferrin as a biomarker of hepatocyte nuclear factor 4 alpha activity and hepatocyte function in liver diseases

doi: 10.1186/s12916-021-01917-6

Figure Lengend Snippet: Visualization of histone modifications by chromatin immunoprecipitation and DNA sequencing (ChIP-seq) in the human transferrin genomic region. ChIP-seq data was obtained from explants from 5 control livers and 7 livers of patients with severe non-responder alcoholic hepatitis. The genomic region around the transferrin promoter was visualized through the Integrative Genome Viewer (IGV). Histone 3 lysine 4 (H3K4) mono- and tri-methylation as well as H3K27 acetylation and tri-methylation are shown. The decrease in H3K27ac signal indicated the lack of activity in transferrin enhancer and promoter regions, while the decrease of H3K4me3 and increase of H3K27me3 suggested the absence of activity and repressive chromatin configuration, respectively, in the region around the transcription start site (TSS). The light gray areas indicate the regions where these changes were seen (cohort ii)

Article Snippet: It was carried out for four histone modifications, using antibodies against histone H3 Lysine 27 acetylation (H3K27ac, Cell Signaling #8173), histone H3 Lysine 27 tri-methylation (H3K27me3, Cell Signaling #9733), histone H3 Lysine 4 mono-methylation (H3K4me1, EDL, Mayo Clinic, Lot#1), and histone H3 Lysine 4 tri-methylation (H3K4me3, EDL, Mayo Clinic, Lot#1).

Techniques: Chromatin Immunoprecipitation, DNA Sequencing, ChIP-sequencing, Methylation, Activity Assay