Fig. 4.

Lymphocyte ChAT expression is transient and reinducible. ChAT-GFP mice were crossed to ChAT.Cre⁺Tomato^LSL mice to allow for fate mapping. Cells from ChAT-GFP⁻ ChAT.Cre⁺Tomato^LSL (Upper) and ChAT-GFP⁺ ChAT.Cre⁺Tomato^LSL (Lower) were assessed by flow cytometry. Tomato⁺ lymphocytes were assessed for GFP in splenic CD4⁺ T cells (A), B220⁺ B cells (B), and peritoneal B1a and B1b B cells (C). (D) Splenic follicular B GFP⁻Tomato⁺ cells were FACS-sorted from ChAT-GFP⁻ ChAT.Cre⁺Tomato^LSL+ and ChAT-GFP⁺ ChAT.Cre⁺Tomato⁺ and stimulated (16 h) with media, LPS (10 μg/mL), or R848 (1 μg/mL). As a control, B220⁺CD23⁺ GFP⁻ cells from ChAT-GFP⁺ ChAT.Cre⁺Tomato⁻ mice were sorted and stimulated. (E) FACS-sorted B2 GFP⁻ or GFP⁺ cells were treated with LPS ± NVP (1 μM) for either 0.5 h or 16 h and were assessed by MS [n = 3 mice per group, representative of three experiments (A–C) and two experiments (D)]. *P < 0.05 by ANOVA; **P < 0.01; ^#P < 0.001.