Fig. 3. Cellular functional changes of etoposide-treated astrocytes in vitro. Scratch wound assay (A–D). The monolayer of rat primary cultured astrocytes were treated with DMSO (vehicle) or 10 μM etoposide for 72 h. (A) Representative images of astrocytes in the wound-healing assay. Blue lines indicate the initial scratch-wound area. Red lines show wound closure at each time-point. Images and data obtained using IncuCyte (Essen Bioscience) under the 10× object. (B) Cell migration every 3 h into the wound area, represented by wound width (μm) in the modified scratch-wound assay analysis. (C) The percentage of relative wound density in the scratch wound area was determined. (D) Etoposide-treated cells were assessed for its phagocytosis ability using E. coli BioParticles. Astrocytes were treated with DMSO (vehicle) or 10 μM etoposide for 12 h. (E) The graph shows total red object area per well (μm2/well) every 10 min for 12 h. (F) The representative graph shows the total red object integrated intensity (RCU × μm2/image) every 10 min for 12 h. Lines represent the mean ± SEM. * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001 vs. vehicle group.
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