Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: The main green tea polyphenol epigallocatechin-3-gallate counteracts semen-mediated enhancement of HIV infection
doi: 10.1073/pnas.0811827106
Figure Lengend Snippet: EGCG targets synthetic PAP248–286-derived amyloid fibrils. (A) Amyloid fibrils were formed by agitation of the fresh PAP248–286 solutions (1 and 5 mg/mL) at 37 °C. Fibrillar aggregates were exposed to various concentrations of EGCG (□, 20 mM; ●, 10 mM; ■, 5 mM; and ▲, 1 mM) and detected by Congo red staining at the indicated time points. Addition of PBS alone (solvent for EGCG) served as negative control (×). (B) Preformed PAP248–286-derived fibrils (SEVI) were treated with increasing concentrations of EGCG (▲, 1 mM; △, 2 mM; □, 4 mM; ○, 8 mM; and ●, 10 mM) for 48 h and analyzed as before. Reactions containing EGCG alone (■, 10 mM) or PBS alone (×) served as controls. (C) HIV-1 particles (X4 strain NL4/3) were preincubated for 20 min with or without the indicated concentrations of SEVI. Subsequently, Jurkat 1G5 luciferase indicator cells were exposed to the respective virus/SEVI mixtures for 5 h. At 24 h after infection, RLU/s was determined. Error bars represent three independent experiments. (D) Cellular metabolic activity was tested in uninfected Jurkat 1G5 cells by alamarBlue assay after 5 h of exposure to the indicated concentrations of SEVI. (E) Viabilities of uninfected Jurkat 1G5 cells were determined after 5 h exposure to the indicated EGCG concentrations as before.
Article Snippet: Synthetic peptides corresponding to PAP (European Molecular Biology Laboratory accession no. {"type":"entrez-protein","attrs":{"text":"AAB60640","term_id":"515997"}} AAB60640 ) amino acid residues 248–286 (PAP248–286) were obtained from Davids Biotechnologie and Bachem.
Techniques: Derivative Assay, Staining, Negative Control, Luciferase, Infection, Activity Assay, Alamar Blue Assay
