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hNT BlueNeurons Express b-Galactosidase at High ,Levels

-Galactosidase Assay Kit. Figure 3 shows beta-galactosidase activity in four representative clones, and their potential for differentiation into neurons was evaluated. Clones were selected according to their ability to proliferate at a sufficient rate and their ability to stably express beta-galactosidase.

beta-galactosidase Expression by hNT Neurons

figure 4

Clones B6, C4 and C6 were chosen to be induced with RA. Cells were cultured in medium containing 10 mg/ml RA for 6 weeks (see Figure 3), and the resulting neurons were harvested and assayed for beta-galactosidase activity as described. As seen in figure 4, clone B6 appeared to have the least activity (about 2 fold above background) but yielded the largest number of neurons among the three clones that were chosen for differentiation.

Figure 5A & 5B

Figure 5A shows in situ staining of control replate 2 cells (after 6 weeks of RA induction, figure 1) that had not been transfected. Neurons were stained with Xgal using Stratagene's In Situ beta-galactosidase Staining Kit. Upon staining, the neurons appear as clusters of small cells that sit on top of accessory cells; a slight blue tinge of background beta-galactosidase activity is apparent. Figure 5B shows transfected B6 clone replate 2 cells. These neurons appear as dark blue small cells in clusters that are easily distinguished from the accessory cell layer. Although both cell types arise from the same transfected precursor cells, the staining of the neurons results in a
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