SRM1649b OF increases IFNγ positive BMDCs in an AHR-dependent manner and generates IL-10 positive BMDCS in an AHR-dependent manner <i>in vitro</i>.

BMDCs were isolated from femurs of WT (C57BL/6J) or Ahr-/- mice. The cells were cultured on bacterial culture plates in complete RPMI media supplemented with GMCSF (10ng/mL) and IL-4 (GMCSF) for 3 days at 37°C and 5% CO2. On day 3, an additional 2.5mLs of media supplemented with GMCSF and IL-4 was added to the culture for an additional 2 days. On day 5, the cells were harvested, resuspended in complete RPMI media, and plated in 96-well plates in the presence of 200ng/mL LPS plus 8 concentrations of SRM1649b OF or solvent control. (A) Representative flow plots of WT IFNγ positive BMDCS, measured by F4/80-CD11c+IFNγ+ cells. (B) Representative flow plots of WT IL-10 positive BMDCS, measured by F4/80-CD11c+IL-10+ cells. (C) SRM1649b OF significantly increased the percent of WT IFNγ positive BMDCs at 10 and 5μg/mL OC (n = 6). (D) There was no significant difference in the percent of IL-10 positive BMDCS between SRM1649b OF and solvent control in WT cells (n = 6) (E) Representative flow plots of Ahr-/- IFNγ positive BMDCS. (F) Representative flow plots of Ahr-/- IL-10 positive BMDCS. (G) In Ahr-/- cells, there was no significant difference in percent of IFNγ positive BMDCS (n = 2) and (H) significant suppression of IL-10 positive BMDCs at 10μg/mL OC (n = 2). Results are mean ± SEM. Significant differences among groups (p<0.05) are indicated by an asterisk.