Figure 2. Rat PC12 cell binding to DI TNC1 astrocytes involved Thy-1/α_vβ₃ integrin interaction.

Fluorescently labelled, live PC12 cells were incubated over a monolayer of DI TNC1 astrocytes. In A) astrocytes in a 24-well plate were treated with 50,100, 300 and 600ng of recombinant Thy-1-Fc proteins of human or mouse origin prior to incubation with labelled PC12 cells. TRAIL-R2-Fc recombinant protein was used as a negative control at 600ng. In B) astrocytes were treated with 5 or 10µg of different anti-integrin antibodies as indicated, for 30min, before the addition of labelled PC12 cells. Treatment with anti-β₁ integrin antibody was used as a negative control in this case. PC12 cells were then incubated for 20min with astrocytes, and subsequently washed gently. Bound labelled cells were counted using an inverted microscope equipped with epifluorescence. Values shown correspond to the percentage of PC12 cells bound to astrocytes whereby non-treated cells (NT) were assigned the value 100%. Averages ± SE of 3 independent experiments are shown. *, p < 0.05 compared to TRAIL-R2-Fc (A) or to anti-β₁ integrin (B) treated cells.