Direct impact of ruxolitinib on T cells and DCs function. (A) ³H thymidine uptake as an indicator of proliferation was measured in mixed lymphocyte reactions containing BALB/c spleen–derived CD4⁺ or CD8⁺ T cells co-cultured with or without C57BL/6 BM-derived DC preactivated with 20 ng/mL LPS. Different concentrations of ruxolitinib were applied. One representative experiment of 3 is shown. (B) ³H thymidine uptake as an indicator of proliferation was measured in mixed lymphocyte reactions containing BALB/c spleen–derived CD4⁺ T cells co-cultured with plate-bound CD3 and soluble CD28 beads. Different concentrations of ruxolitinib were applied. One representative experiment of 2 is shown. (C) Viability of CD4⁺ T cells derived from cultures described under (A) are shown. Live cells were determined by the number of viable cells reacting with the dye used in the live/dead stain kit. One representative experiment of 3 is shown. (D) Intracellular Granzyme B production in CD8⁺ T cells derived from cultures described under (A) are shown. (E) Proinflammatory cytokines measured in the supernatants of cultures described under (A) are shown. One representative experiment of 3 is shown. (F-G) STAT3 phosphorylation as determined by phospho flow of CD4⁺ T cells derived from cultures described under (A) are shown. One representative experiment of 3 is shown.