Fig. 4.

Microglia aggravated free heme-induced neuronal apoptosis. A BV2 cell and HT22 cell coculture system and four groups: 1) Neuron group (HT22) in which only HT22 cells were incubated for 24 h; 2) Neuron Heme group (HT22 + Heme) in which HT22 cells were treated with hemin for 24 h; 3) Neuron + microglia group (HT22 + BV2) in which HT22 cells cocultured with BV2 cells were incubated for 24 h; 4) Neuron + microglia Heme group (HT22 + BV2 + Heme) in which HT22 cells cocultured with BV2 cells were treated with hemin for 24 h. B Representative FACS plots of apoptotic cells (Q2 + Q4) in four groups. C Quantification of the percentage of apoptotic cells among HT22 cells in four groups. D Representative immunofluorescence images of nuclei (blue) and NeuN (red) staining among HT22 cells in four groups. Scale bar = 100 μm. E Quantitative analysis of NeuN + cells in four groups. Data are presented as means ± SEM. ***p < 0.001. *p < 0.05