Identification of the Ank1 and Ank6 domains that are essential for the ability to co-immunoprecipitate p65, importin β1, and exportin 1. Sean M. Evans Kyle G. Rodino Haley E. Adcox Jason A. Carlyon 10.1371/journal.ppat.1007023.g015 https://plos.figshare.com/articles/figure/Identification_of_the_Ank1_and_Ank6_domains_that_are_essential_for_the_ability_to_co-immunoprecipitate_p65_importin_1_and_exportin_1_/6228755 <p>Whole cell lysates of HeLa cells expressing the indicated Flag-tagged proteins were incubated with Flag antibody-conjugated agarose beads to immunoprecipitate Flag-tagged proteins and their interacting proteins. (A) Western blots of input lysates and immunoprecipitates were probed with p65, exportin 1 (Exp 1), and importin β1 (Imp β1) antibodies. Expression of each Flag-tagged protein of interest was confirmed by subjecting input lysate to Western blotting using Flag antibody. GAPDH immunoblotting of input lysates served as a loading control. (B) Mean <u>+</u> SD of co-immunoprecipitated p65, exportin 1, or importin β1. Values are presented as the percent change in densitometric signal relative that for each respective protein of interest that was coimmunoprecipitated by Flag-tagged Ank1 or Ank6. Data presented were calculated from three separate experiments formed in (A). Statistically significant (*<i>P</i> < 0.05; **<i>P</i> < 0.01) values are indicated.</p> 2018-05-07 17:37:21 blocks importin β1 activity Flag-tagged Ank 1 exploits host exportin 1 p 65 NF -κB transcription NF -κB transcriptional activation Orientia tsutsugamushi causes abrogates Ank 1 exportin 1 tsutsugamushi antagonizes NF -κB transcription Ank 1 ankyrin repeat-containing effectors bind importin β1 antimicrobial response Ank 6 co-immunoprecipitate p 65 NF -κB NF -κB p 65 TNF α-induced p 65 Ank 6 regions bacterium modulates p 65 subcellular localization exportin 1- Ank 6 F-box