Fig. 4

Response to excitotoxicity. a Representative fluorescence images of the co- and tri-cultures at 48 h following exposure to 25 μM glutamate for 1 h. The cultures were immunostained for the three cell types of interest: neurons—anti-βIII-tubulin (red), astrocytes—anti-GFAP (green), microglia—anti-Iba1 (orange), and the general nuclear stain DAPI (blue). Scale bar = 100 μM. Representative images from the other conditions can be found in Supplementary Figure 4. b Comparing the average astrocyte area between the co- and tri-cultures following challenges with different concentrations of glutamate. A full analysis of the simple main effects can be found in supplementary data table 1. c The neuron percent area coverage, with a 0.2 circularity cutoff to eliminate cell debris, following excitotoxic challenge. A full analysis of the simple main effects can be found in supplementary data table 2. d The average microglia surface area did not change following treatment with different concentrations of glutamate. e Calcium imaging results showing the change in fluorescence intensity following treatment with different concentrations of glutamate. A full analysis of the simple main effects can be found in supplementary data table 3. b–e All graphs display mean ± SD (n = 3), while the points indicate the values of the technical replicates. The letters above the bars indicate statistically significant differences (p < 0.05)