NleC cleaves p65 during <i>C</i>. <i>rodentium</i> and EPEC infections.

<p><b>A</b>. HEK293T cells expressing N-terminal GFP-tagged p65 were mock infected or infected with indicated <i>C</i>. <i>rodentium</i> strains at 100 MOI for 3 h. Whole cell lysates were derived, separated by SDS/PAGE, and immunoblotted for indicated proteins. The GFP-tagged p65^1–10 cleavage products are indicated by an arrow, and an asterisk labels nonspecific bands. <b>B</b>. Caco-2 cells were mock infected or infected with indicated EPEC strains at 100 MOI for 1 h. Whole cell lysates were derived, separated by SDS/PAGE, and immunoblotted for indicated proteins. <b>C</b>. Mouse colon epithelial cells (CECs) were mock infected or infected in suspension with indicated <i>C</i>. <i>rodentium</i> strains at 100 MOI for 3 h. Whole cell lysates were derived, separated by SDS/PAGE, and immunoblotted for indicated proteins. The full-length p65 and cleaved p65 C-terminal fragment are indicated by filled and open triangles, respectively. <b>D</b>. Mouse CECs infected as in C were separated from free <i>C</i>. <i>rodentium</i> by Percoll gradient centrifugation. Whole cell lysates from CECs and attached <i>C</i>. <i>rodentium</i> were derived, separated by SDS/PAGE, and immunoblotted for indicated proteins. <b>E</b>. Representative immunofluorescence micrographs of mouse CECs infected as in C that were centrifuged onto cover slips and stained for <i>C</i>. <i>rodentium</i> LPS, with nuclei counterstained by DAPI. Scale bars, 10 μm. <b>F</b>. The numbers of <i>C</i>. <i>rodentium</i> attached to mouse CECs as in E (from 6 random fields) were quantified and normalized to the perimeter of individual CEC.