Cytokine production, proliferation and maturation of BMDCs after stimulation with LPSpa.

<p>BMDCs were obtained from C57BL/6 mice-derived bone marrow cells cultured with GM-CSF. Cells were stimulated for 24 h with MPL (0.1, 1, 10, 100, 1000 ng/ml) or LPSpa (0.1, 1, 10, 100, 1000 ng/ml) and (A) IL-6 or (B) TNF levels in supernatants were measured by ELISA (n = 3). (C) Total cell numbers were counted (n = 3) and (D) the maturation of BMDCs was assessed by measuring the expression levels of CD40, CD80 and CD86 by flow cytometry (n = 3). (E) Cells were stimulated for 8 h with LPSpa (1000 ng/ml) and then stained with propidium iodide and analyzed by flow cytometry (n = 3). Results are representative of three (C, D), four (E) or five (A, B) independent experiments. <i>*P</i><0.05, <i>**P</i><0.01, <i>***P</i><0.001, by Two-tailed student <i>t</i>-test when comparing between two groups and one-way ANOVA with Dunnett’s test when comparing three or more groups.