Fig. S6.

Noncognate iNKT cells increase in number but do not preferentially expand a greater percentage of IL-10⁺ B cells. FACS analysis was used to identify NP-specific B10 (B220⁺CD19⁺CD1d⁺CD5⁺IL-10⁺) cell numbers 4 d after immunization of C57BL/6 mice with vehicle (PBS/0.1% BSA), 100 μg NP-KLH plus 0.5 μg αGalCer (αGC), or 0.5 μg NPαGalCer (NPαGC). FACS analysis specifically revealed (A and B) B220⁺NP⁺, (C and D) B220⁺NP⁺CD1d⁺CD5⁺, and (E and F) B220⁺NP⁺CD1d⁺CD5⁺IL-10⁺ (one experiment; n = 3–5 mice per group;). *P ≤ 0.05. (G) VertX IL-10 reporter mice were immunized i.v. with vehicle (PBS/0.1% BSA), 100 μg NP-KLH plus 0.5 μg αGC, 0.5 μg NPαGC, or LPS, and their splenocytes were assessed 4 d later by FACS analysis to determine the percentage of B10 cells (B220⁺CD19⁺CD1d⁺CD5⁺IL-10⁺) within the expanded antigen-specific CD5⁺ CD1d⁺ B regulatory cell population (representative of three experiments; n = 3–5 mice per group).