Fig. 3

Chronic IFNγ treatment increases PDGFRβ phosphorylation and internalization. a Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh) or IFNγ (1 ng/mL). After 96 h total treatment, cells were serum starved for 2 h and then treated with vehicle (−) or PDGF-BB (100 ng/mL) for 30 min as depicted. b Representative western blots of treated pericytes (n = 3). c–e Bands were quantified with Image Studio™ and normalized to vehicle control. p-PDGFRβ (c) was normalized to total PDGFRβ, and p-Akt (d) and p-ERK (e) were normalized to total Akt and ERK, respectively. f Representative images of pericytes treated as in a showing PDGFRβ (green) and Hoechst (blue), scale bar, 10 μm. g PDGFRβ puncta in f were quantified using MetaXpress™ software and normalized to cell number and vehicle control and plotted as mean ± s.e.m. (n = 3), ****(p < 0.0001), ^###(p < 0.001), *(p < 0.05) (two-way ANOVA). h Cellular localization of PDGFRβ (green) and nuclei (blue) in pericytes treated with vehicle (Veh) or PDGF-BB (100 ng/mL) for 30 min as above using confocal microscopy. Scale bar, 5 μm