Fig. 1

Chronic IFNγ treatment alters αSMA, but not PDGFRβ expression, or cell number in human brain pericytes. a Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh) or IFNγ (1 ng/mL) as depicted. b, c Cells were then fixed and imaged under brightfield (b), and total cells counted from Hoechst labelled nuclei (c). d–g Representative images and quantification of PDGFRβ (green) (d, e), or αSMA (red) (f, g) overlaid with Hoechst. Scale bar, 100 μm. The integrated intensity of the staining was normalized to cell number (Hoechst) (c) and vehicle conditions, quantified from triplicate wells, and plotted as mean ± s.e.m. (n = 3) and ****(p < 0.0001) (Student’s t test). h, i mRNA from pericytes treated as in a was analysed by qRT-PCR. Inflammatory target gene (IP-10, MCP-1, COX2, ICAM-1, CD74) (h) and pericyte marker and proliferation marker gene (PDGFRβ, αSMA and Ki67) (i) expression was normalized to GAPDH and plotted as a mean fold change from vehicle (set to 1) (2^ΔΔCT) ± s.e.m. (n = 3), **(p < 0.01) by a Mann-Whitney, non-parametric test of ΔCT values