Fig. 5

MJ04 inhibits the differentiation of naïve T-cell into Th1 and Tc1 subsets, also inhibits the level of proinflammatory cytokines in LPS primed macrophages. Splenoctyes from healthy mice were used as a source of T-cells, Spelnocytes were stimulated with anti-CD3/C28 in the presence and absence of MJ04 (0.01, 0.1, 1 and 10 µM) for 96 h and before harvesting these cells were further stimulated with PMA/Ionomycin for additional 4 h. A–D is the gating strategy to deduce the cell of interest. The expression of IFN-γ in the CD4⁺(C, D) and CD8⁺ (E, F) T-cells were analyzed by flowcytometry. In another set of experiments, mouse macrophage (J774a.1) were induced with LPS and then treated with different concentration of MJ04 and Tofacitinib. The level of pro inflammatory cytokines (TNF-α, IL1-β and IL-6) were measured by sandwich ELISA from LPS induced macrophages (G–I). Each data point is representative of n = 3). Data points are represented as mean ± SEM and value of significance represents *P < 0.05, **P < 0.005, ***P < 0.0005 and ****P < 0.00005