GSK-3 inhibition after TLR4 ligation regulates expression of IL-12 in CD86^lo DCs. Murine CTR- and RAPA-DCs were treated for 2 hours with an Akt (AktX; 5μM) or GSK-3 (LiCl; 10mM) inhibitor before 18-hour exposure to 0.1 μg/mL LPS. (A) Staining for CD86 and IL-12 confirmed that increased IL-12 could be ascribed to a population of CD86^lo RAPA-DCs. These analyses also demonstrated that inhibition of GSK-3, but not AKT, limited IL-12p40 expression in CD86^lo DCs, especially RAPA-DCs. (B-C) Comparison of the percentages (mean ± SD) of IL-12p40⁺CD11c⁺ cells (B) between groups demonstrated a significant decrease in LiCl-treated CTR and RAPA-DCs. (C) However, on more detailed analysis, a significant difference between RAPA- and CTR-DCs was only observed in CD86^lo, not CD86^hi, mDCs. CD86^lo DCs were the only subset that displayed a significant decrease in IL-12p40 after inhibition of GSK (n = 2-4). *P < .05.