Figure 7

FPRs-induced neuronal differentiation of NSCs was mediated by ROS generation. (A–F) FPR1-induced neuronal differentiation of NSCs was mediated by ROS generation. (A,B) Flow cytometric analysis for generation of ROS by NSCs incubated in 0, 0.5, 1, and 5 µM fMLF for 6 h with or without preadministering NAC at 5 mM. (A) Representative plots of FPR1-mediated ROS generation. (B) Histograms for quantification of ROS generation. The levels of ROS were represented by the median fluorescence intensity. (C,D) Dissociated NSCs were differentiated with or without preadministering NAC for 30 min before administering fMLF. The images in (C) are representative of several independent experiments, and the graph in (D) shows the statistical results. (E) Dissociated NSCs were differentiated in an extremely low density (1000 cells/mL), and then the cells were immunostained with β-III tubulin. (F) Quantified analysis of the total length of neurites per cell, the number of primary neurites, and the number of branch points. (G–L) FPR2-induced neuronal differentiation of NSCs was mediated by ROS generation. (G,H) Flow cytometric analysis for generation of ROS by NSCs incubated in 0, 0.05, 0.1, and 0.5 µM MMK-1 for 6 h with or without preadministering NAC at 5 mM. (G) Representative plots of FPR1-mediated ROS generation. (H) Histograms for quantification of ROS generation. (I,J) Dissociated NSCs were differentiated with or without preadministering NAC for 30 min before administering MMK-1. The images in (I) are representative of several independent experiments, and the graph in (J) shows the statistical results. (K) Representative micrographs of the differentiated cells immunostained with β-III tubulin. (L) Quantified analysis of the total length of neurites per cell, the number of primary neurites, and the number of branch points. Data were presented as mean ± SEM; *P < 0.05; **P < 0.01; N = 4. Scale bar = 20 µm.