Fig. 9

SCF + G-CSF treatment enhances microglia in engulfing synaptosomes in vitro. a Western blotting shows that isolated synaptosomes express synaptic proteins. b A schematic flowchart of the in vitro study. c Representative western blot images show that PSD-95 and Gephyrin are expressed in the cultured microglia after adding synaptosomes. d and e Quantification data of western blots show that the levels of PSD-95 (d) and Gephyrin (e) in the cultured microglia are increased by SCF + G-CSF treatment. Control: n = 4, SCF + G-CSF treatment: n = 4. f Representative immunocytochemistry images show the engulfed DiO-labeled synaptosomes (green) in cultured microglia (red, CX3CR1 positive). g The scatterplot bar graph shows that SCF + G-CSF treatment increases uptake of synaptosomes (DiO positive area) by the cultured microglia. Each dot represents a single microglial cell. Control: n = 60 microglia, SCF + G-CSF treatment: n = 60 microglia. Data were collected from four independent experiments. h The cumulative frequency distribution of DiO positive area in microglial cells. Note that the engulfed synaptosomes (DiO positive area) by the cultured microglia are increased by SCF + G-CSF treatment. Kolmogorov–Smirnov test. i Representative scatterplot graphs of flow cytometry show the strategy of data analysis. The doublets and debris are excluded, and only the DiO positive cells are analyzed. j The bar graph shows the DiO fluorescence density in microglial cells. k The bar graph shows the percentage of DiO positive microglial cells. Control: n = 3, SCF + G-CSF treatment: n = 3. Student’s t test. Mean ± SEM. *p < 0.05, ****p < 0.0001