Enteric glial MHC-II signaling contributes to IFNγ/LPS-induced immunocyte activation in mesenteric lymph nodes. (A) Flow cytometry gating strategy to select cells based on the appropriate forward scatter (FSC) and side scatter (SSC) (left); single events based on the FSC area (FSC-A) and height (FSC-H) (middle); and live cells without the Zombie Yellow viability dye (right). (B) Representative data of an early activation marker CD69 expression in cells from the mesenteric (top row) and inguinal lymph nodes (bottom row) that were harvested from mice treated with IFNγ and saline (control are pooled glialMHC2KO animals and wild-type littermates, left column), IFNγ- and LPS-treated wild-type littermates (WT + LPS, middle column), and IFNγ- and LPS-treated glialMHC2KO mice (WT + knockout [KO], left column). (C) Percentage of single live cells expressing CD69 in the mesenteric and inguinal lymph nodes of glialMHC2KO animals (KO) and wild-type littermates (WT) before (controls) and after LPS stimulation (n = 4–6 mice); 2-way analysis of variance with Sidak’s-corrected multiple comparisons; nsP = .424, ∗P = .022, and ∗∗∗∗P < .0001. LN, lymph node.