Fig. 3

Deleterious effects of factors released by 22L-astrocytes on neuronal morphology, viability and expression of functional genes. a Analysis of expression of synaptogenic and neurotrophic genes in 22L-PACs normalized by the expression levels in CT-PACs using qRT-PCR. b Schematic illustration of experiments on treatment of mouse primary cortical neuronal cultures with astrocyte-conditioned media (ACM) collected from CT-PACs and 22L-PACs. c Representative images of primary neuronal cultures isolated from P1–P2 mice and co-immunostained for MAP2 (green), NeuN (red) and DAPI (blue). d Left: representative images of primary neuronal cultures treated with CT-ACM and 22L-ACM for 72 h and co-immunostained for MAP2 (green) and NeuN (red). Right: quantification of neuronal morphology using MAP2 fluorescence in primary neuronal cultures treated with CT-ACM and 22L-ACM for 72 h. e Analysis of expression of Syp, Syn2, Dlg4, Thbs2, Gria1 and Gria4 in primary neuronal cultures treated with 22L-ACM for 72 h and normalized by the expression levels in cultures treated with CT-ACM using qRT-PCR. f Cell viability in primary neuronal cultures assessed by MTT assay as a function of incubation time with CT-ACM or 22L-ACM. In panels a and e, Gapdh was used as housekeeping gene. In panels. a, d–f, data represent means ± SE, n = 3 independent astrocyte or microglia cultures isolated from individual animals, ***p < 0.001, **p < 0.01, *p < 0.05, and ‘ns’ non-significant (two tailed, unpaired student t test). Scale bar = 50 µm