Schulz, Danae R. Mugnier, Monica Paulsen, Eda-Margaret Kim, Hee-Sook W. Chung, Chun-wa F. Tough, David Rioja, Inmaculada Prinjha, Rab K. Nina Papavasiliou, F. Debler, Erik W. Bromodomain proteins are necessary for BF-specific antibody internalization. <p>(A) Flow cytometry measuring the amount of primary anti-VSG antibody remaining on the surface following a 5-min incubation at the indicated temperatures, washout, fixation, and incubation with secondary antibody in I-BET151-treated (right) versus control cells (left). "Secondary" (gray trace) denotes control cells stained with secondary antibody only. (B) Immunofluorescence showing the amount of primary anti-VSG antibody remaining on the surface following a 5-min incubation at the indicated temperatures, washout, fixation, and incubation with secondary antibody in I-BET151-treated versus control cells. A bright spot is detected at the flagellar pocket in control cells incubated at 4°C, whereas I-BET151-treated cells have antibody distributed over the entire surface. At 37°C, control cells have internalized almost all the primary antibody. (C) Flow cytometry showing the amount of anti-VSG3 antibody bound to the surface in cells treated with I-BET151 for 3 d, compared to control cells. (D) Anti-VSG Western blot showing that protein levels of <i>VSG</i> at the active expression site (ES) are unaltered in I-BET151-treated cells. Tubulin is used as a loading control. Numerical data for Fig 3 is in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002316#pbio.1002316.s003" target="_blank">S3 Data</a>.</p> Bloodstream Form Stage Identity;novel binding mode;immunogenic surface VSG;African Trypanosome Trypanosoma brucei;bromodomain inhibition;host antibody responses;1.25Å resolution crystal structure;BF cell fate;bromodomain proteins;trypanosome bromodomain proteins;bloodstream stage identity;immunogenic Variant Surface Glycoprotein 2015-12-08
    https://plos.figshare.com/articles/dataset/_Bromodomain_proteins_are_necessary_for_BF_specific_antibody_internalization_/1621516
10.1371/journal.pbio.1002316.g003