Fig. 1

Astrocytes isolated from 22L-infected mice exhibit a reactive phenotype. a Analysis of gene expression in 22L-PACs, normalized by the expression of the same genes in CT-PACs, using qRT-PCR. b Representative Western blots and densitometric analysis of GFAP expression normalized per expression of β-actin in CT-PACs and 22L-PACs. c Representative images of CT-PACs and 22L-PACs stained for GFAP, and morphometric analyses of cell area, perimeter and process number in astrocytes from CT-PACs and 22L-PACs. Insets show magnified images. d Analysis of expression of PAN-, A1- and A2-specific genes in 22L-PACs normalized by the expression levels in CT-PACs using qRT-PCR. e Representative images of co-immunostaining of CT-PACs and 22L-PACs for GFAP (green), LCN2 (red) and nuclei (DAPI, blue), and quantification of integrated fluorescence intensity of LCN2 in CT-PACs and 22L-PACs. f Representative Western blots and densitometric analysis of LCN2 expression normalized per expression of β-actin in CT-PACs and 22L-PACs. g Analysis of expression of pro-inflammatory genes in 22L-PACs normalized by the expression levels in CT-PACs using qRT-PCR. h Analysis of IL-6 concentration in media conditioned by CT-PACs and 22L-PACs. In panels a, d and g, Gapdh was used as a housekeeping gene. In panels a–h, data represent means ± SE, n = 3 independent cultures isolated from individual animals, ***p < 0.001, **p < 0.01, *p < 0.05, and ‘ns’ non-significant (two tailed, unpaired t test). Scale bar = 50 µm for panels c and e