Primary human neuron-glia cultures exposed to RAL. (A) Quantification of area of β-III-tubulin (average area, μm2), and GFAP in neuron - glia cultures exposed to 0, 20, and 100 nM RAL for 15 hr. Representative images of cultures treated with (B) Control, (C) 20 nM, and (D) 100 nM RAL. Cultures were maintained for four weeks in neurobasal media on glass coverslips then exposed to RAL overnight, cells were fixed and stained for β-III tubulin (green) and GFAP (red) to illustrate neurons and astrocytes, respectively. Ten images were acquired by deconvolution fluorescence microscopy under 40X objective magnification and quantified using Image Pro Plus.