Fig. 3

Astrocytes from P301S mice have a reduced capacity to support neuronal survival. Primary astrocytes (C57A and P301SA) cultured from cerebral cortex of 7 day-old mice (98% purity) were plated on top of primary neurons cultured from mice of similar age and brain region for 4–5 days. Co-cultures were maintained for 4 and 8 days. a Representative images of co-cultures immunostained for β-III-tubulin (red), GFAP (green) and Dapi (blue). Quantification of neuron (b, c) and astrocyte (d, e) numbers after 4 and 8 days of co-culture. Each experiment consisted of six technical replicates (wells) in which at least five fields were analyzed. Data show mean per field ± SEM from at least four independent experiments. Data were analysed using ANOVA followed by Tukey’s multiple comparison test; *p < 0.05 for these comparisons: C57N vs C57N + P301SA; C57N vs P301SN + P301SA; C57N vs C57N + C57A; C57N + C57A vs P301SN + C57A; C57N + C57A vs C57N + P301SA; C57N + C57A vs P301SN + P301SA; P301SN + C57A vs P301SN + P301SA; ANOVA of results from 4 day co-cultures revealed a significant interaction between genotype and co-culture condition [F (2, 21) = 4.477; p = 0.0240], significant effects of co-culture type [F (2, 21) = 14.27; p = 0.0001] and genotype [F (1, 21) = 14.8; p = 0.0009]. In 8 day of co-cultures, ANOVA revealed no interaction between genotype and culture conditions [F (2, 22) = 3.048; p = 0.0678], the significant effect being co-culture type [F (2, 22) = 17.51; p < 0.0001] and co-culture condition [F (1, 22) = 6.54; p = 0.0180]. No significant differences in the numbers of astrocytes were present between the various cultures