Fig. 7

Chronic IFNγ treatment reduces PDGFRβ re-synthesis following ligand-stimulated degradation. a–c Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh) or IFNγ (1 ng/mL); the final 48 h was in the presence of either vehicle or PDGF-BB (10 ng/mL), with representative images of PDGFRβ (green), αSMA (red), and Hoechst (blue) (a) Scale bar, 100 μm. Quantification of PDGFRβ (b) and αSMA (c) staining, mean ± s.e.m. (n = 3), ^####, ****(p < 0.0001), ***(p < 0.001), **(p < 0.01) (two-way ANOVA). d–f Pericytes were treated for three or four consecutive days (once every 24 h) with either vehicle (Veh) or IFNγ (1 ng/mL). After 48 h, cells were treated with PDGF-BB (10 ng/mL) for either 24 or 48 h. Representative blots of PDGFRβ, αSMA, and GAPDH (d) Quantification of band intensity of PDGFRβ (e) and αSMA (f), both normalized to GAPDH, mean ± s.e.m. (n = 3), **(p < 0.01), *(p < 0.05) (two-way ANOVA). g–i Pericytes were pre-treated for 48 h (once every 24 h) with either vehicle or IFNγ (1 ng/mL) and then treated with PDGF-BB (10 ng/mL) for 24, 48, 72, or 96 h (with IFNγ being added every 24 h throughout). qRT-PCR analysis of PDGFRβ (g), αSMA (h), and Ki67 (i) transcripts was normalized to vehicle treatment and plotted as mean ± s.e.m. (n = 3), ^####, ****(p < 0.0001), ^##, **(p < 0.01), *(p < 0.05) (two-way ANOVA)