Fig. 5
TGFβ1 attenuates phagocytosis by pericytes. Pericytes were treated for 24 h with 0–10 ng/mL TGFβ1 in 1 mM citric acid, pH 3 with 0.1 % BSA, followed by a further 24 h in the presence or absence of a 1:1000 dilution of fluorescent latex beads. Cells were fixed and nuclei counterstained with DRAQ5. Representative images for vehicle and 10 ng/mL TGFβ1 treatments are shown (a). The percentage of phagocytic pericytes was determined by automated image analysis from five independent experiments (b). Confocal microscopy of pericytes plated on glass coverslips, incubated for 24 h with a 1:10,000 dilution of fluorescent beads and immunostained with PDGFRβ confirmed internalisation of beads (c). Pericytes were treated for 24 h with 0–10 ng/mL TGFβ1 in 1 mM citric acid, pH 3 with 0.1 % BSA. For an additional two hours cells were incubated in the presence or absence of a 1:1000 dilution of fluorescent latex beads. Phagocytosis was determined by flow cytometry. One representative plot is shown (d) and the mean fluorescent intensity (MFI) of five independent experiments was determined (e). *p < 0.05, **p < 0.01, ***p < 0.001 versus vehicle control (ANOVA). Scale bar = 50 μm