Fig. 5

22L-derived reactive microglia induce a pro-inflammatory state in primary astrocytes. a Left: representative images of primary astrocyte cultures isolated from adult (200–300 days old) C57 Black mice, treated with CT-MCM or 22L-MCM for 72 h, and co-immunostained for GFAP (green) and nuclei (DAPI, blue). Right: morphometric analysis of astrocytes following treatment of primary cultures with CT-MCM or 22L-MCM and staining for GFAP. Data represent means ± SE, n = 3 independent experiments, **p < 0.01 and *p < 0.05 (two tailed, unpaired student t test with Welch's correction). Scale bar = 50 µm. b and c. Analysis of expression of PAN-, A1- and A2-specific genes (b) and synaptogenic genes (c) in primary astrocyte cultures following treatment with CT-MCM or 22L-MCM for 72 h using qRT-PCR. Gapdh was used as housekeeping gene. Data represent means ± SE, n = 3 independent experiments, i.e. astrocyte cultures isolated from individual animals was treated by CT-ACM or 22L-ACM collected from cultures also established form individual animals, ***p < 0.001, **p < 0.01 and *p < 0.05 and ‘ns’ non-significant (two tailed, unpaired student t test)