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. Author manuscript; available in PMC: 2012 May 1.
Published in final edited form as: Stem Cell Res. 2011 Jan 19;6(3):195–205. doi: 10.1016/j.scr.2011.01.002

Figure 1. Characterization of ES-derived motoneurons.

Figure 1

Figure 1

(A) Western blot analyses of cellular markers expressed at different stages of the differentiation process. Markers included eGFP driven by Hb9 motoneuron promotor (Hb9-eGFP) genes expressed by pluripotent ES cells, Oct4 and Sox2; neuronal cells, Tuj1 and Tau; and post-mitotic motoneurons, ChAT protein. Similarly, SNAP-25 protein expression, the only-known intracellular target of BoNT/A, was also evaluated. Total protein loaded per lane was 10–15 µg and GAPDH was used as loading control. (B) Immunostaining for neural markers in the differentiated cultures. Cells from embryoid bodies dissociated at day 7 and cultured on matrigel-coated culture slides for an additional 3 days were immunostained with antibodies against neuronal marker, NeUN; post-mitotic motoneuron markers Hb9, and Isl1; SNARE protein, SNAP-25, and SV2A, the receptor required for BoNT/A entry into neurons. Shown are also the Hb9-eGFP and DAPI nuclear staining. The scale bar is 50 µm.