FIG. 7.

S63845 treatment reduces the intrahepatic B lymphocyte population and does not deplete the systemic pool of B lymphocytes. Intrahepatic leukocytes from DKO mice treated with vehicle or S63845 (40 mg/kg) daily for 5 days were stained with a panel of 24 cell surface markers and analyzed by CyTOF (n = 3). (A) Twenty-nine distinct clusters of intrahepatic leukocytes are visualized on a t-distributed stochastic neighbor embedding plot. (B) Representative density plots of vehicle and S63845-treated groups displaying the change in clusters. Red indicates high frequency categorization of cells to a cluster; blue indicates low frequency. (C) Distribution of B cells across clusters which were identified by CD45⁺B220⁺CD19⁺MHCIIhi (B lymphocytes) and CD45⁺B220+CD19⁺Tim4⁺MHCIIhi (a proinflammatory B-cell cluster). (D) Seven clusters that identified B cells, including cluster 18, which was specific for the proinflammatory phenotype. (E) DKO mice were treated with vehicle or S63845 (40 mg/kg) daily for 5 days, and spleens were harvested. Representative fluorescence-activated cell sorting analysis of primary splenic cells showing single CD45⁺B220⁺ cells (B lymphocytes). Graph represents the percentage of B lymphocytes (n = 3). Data are expressed as mean ± SEM. *P < 0.05, **P < 0.01. Abbreviations: APC, allophycocyanin; ns, not significant; PERCP, peridinin chlorophyll protein; tSNE, t-distributed stochastic neighbor embedding.