Fig. 5
Iron is essential only for the middle and late stages of BCR- and TLR-induced B-cell proliferation. a Intracellular calcium flux in control and iron-deficient (pretreated with 20 µM DFO for 2 h) resting B cells purified from wild-type C57BL/6 splenocytes in response to BCR stimulation. b Control and iron-deficient (pretreated with 20 µM DFO for 2 h) B cells were stimulated with anti-IgM (10 µg/ml) for 0–20 min. At the indicated time points, the cells were immediately collected, washed, and lysed. Subsequently, the phosphorylation of ERK1/2 was measured by immunoblotting. c CFSE-labeled splenic B cells from wild-type mice were unstimulated or stimulated with anti-IgM and LPS, and cell proliferation was assessed by CFSE dilution at 72 h. During this process, DFO (20 µM) was added to the culture medium at different time points to deplete iron. d CFSE-labeled iron-deficient B cells (in the presence of 20 µM DFO) were stimulated as in c, and cell proliferation was assessed at 72 h. During this process, FAC was added to the culture medium at different time points to restore the iron supply. The data are representative of three independent experiments
