Fig. 7
Culture medium does not alter pericyte responsiveness to growth factors TGFβ1 and PDGF-BB. Human brain pericytes were plated at a low density and allowed to proliferate in either DMEM/F12 with 10% FBS or Pericyte Medium for 7 days. For the final 2 h pericytes were incubated with 10 ng/mL TGFβ1 or vehicle (1 µM citric acid, pH 3 with 0.0001% BSA). Cells were fixed and immunostained for SMAD2/3 (a) and the nuclear intensity was determined by automated imagining analysis (b). Human brain pericytes were plated at a low density and allowed to proliferate in either DMEM/F12 with 10% FBS or Pericyte Medium for 7 days. For the final 30 min pericytes were treated with 10–100 ng/mL PDGF-BB or vehicle (4 µM hydrochloric acid with 0.0001% BSA). Cells were brought into suspension with Accutase and incubated with a PDGFRβ antibody or an isotype control. Cell surface PDGFRβ was determined by flow cytometry (c). Data are displayed as mean ± SEM of three independent experiments. ***p < 0.001 versus vehicle control in respective media (Two-way ANOVA). Scale bar = 50 µm