Statistical data showing the effect of AgNPs on the mean fluorescent intensity of cytoskeletal proteins and representative images showing the impact of AgNPs on glial β-tubulin and GFAP. To quantify the effect of AgNPs on the level of β-tubulin and F-actin, the fluorescent intensity of randomly selected 7-16 areas of 3844 μm2 under control and AgNP-treated conditions (as shown in Figure 4F-4J) were measured using imageJ software. The quantified data show that AgNPs significantly decreased the fluorescent intensity of both β-tubulin (A) and F-actin (B). To further investigate whether AgNPs affected glial cytoskeleton, cells were co-stained with β-tubulin (green) and GFAP (red), a specific marker for glial astrocytes. C shows the representative images of cells in the control and AgNPs at 10 μg/ml. Neurons under the control conditions exhibited healthy neuronal cell bodies (indicated by a solid arrow) and neurites (indicated by an open arrow), while neurons exposed to 10 μg/ml of AgNPs had degenerated cell bodies (indicated by a solid arrow) and neurites (indicated by an open arrow). Note that both of the β-tubulin and GFAP in glial cells (indicated by asterisks) were compromised by AgNPs. * P < 0.05. ** P < 0.01. *** P < 0.001. Error bars indicate SEM. Scale bar, 20 μm.