Clinical concentrations of CsA do not induce apoptosis of murine BMDCs. On day 3, DCs cultured in GM-CSF and IL-4 were treated with different concentrations of CsA, as indicated, for 72 hours. On day 6, DCs were harvested and analyzed. (A) Apoptosis assay of CsA-treated DCs. DCs treated with CsA were labeled with annexin V/PI (left; numbers indicate percentages of PI- or annexin V–positive cells), DHR123 (middle; labels indicate mean fluorescence intensity of shaded histogram), and rhodamine 123 (rho123; right; labels indicate percentage of R123^low DCs) to detect early apoptosis, ROS production, and mitochondrial membrane potential, respectively. (B) Western blot analysis of apoptosis-associated molecules in DC lysates.