Fig. 6

miR-574-5p exposure leads to potent activation of human and mouse B lymphocytes. NS, not significant; ** P < 0.01, *** P < 0.001. (a) miR-574-5p exposure in human PBMCs significantly increased both total B cells (CD19⁺) and activated B cells (CD19⁺CD69⁺) (n = 3). About 1 × 10⁶ human PBMCs were seeded in 6-well plate and treated with 1 µg/ml R848 or 10 µg/ml of Dotap-PS-miR-574-5p for 24 h. Subsequently, the cells were harvested for flow cytometry. (b) miR-574-5p exposure potently stimulated the activation of human PBMC-derived B cells (n = 4–5). B cells were purified from human PBMCs of three normal healthy subjects and treated as human PBMCs as described above. (c) miR-574-5p exposure significantly increased splenic B cell activation in the B6.WT mice but not the B6.Tlr7^−/− mice. About 1 × 10⁵ splenocytes were seeded in 6-well plates and treated with 10 µg/ml of Dotap-PS-miR-574-5p or 1 µg/ml of R848 for 24 h. B cell activation was analyzed by flow cytometry gated on B220⁺Cd69⁺ (n = 4)