Chapter 2 76 Supplementary Figure 2. Pilot study to test efficacy of SAG concentrations. A control and a 4H line were treated with vehicle, 3 nM, 50 nM, 100 nM or 250 nM SAG, either started from day 18 or day 30 of the neuronal differentiation. (A) The left graph shows the percentage of VGAT+ synapses, which did not change in the control line in any treatment condition. However, for the 4H line the percentage of VGAT+ synapses was increased after treatment with 100 or 250 nM SAG. The right graph shows the number of synapses per dendritic length. SAG treatment decreased the number of synapses per dendritic length slightly in the control line, although it was slightly increased in the 4H line with 50 or 100 nM SAG when treatment was started at day 18. (B) shows the expression of SHH downstream target GLI1 and housekeeping gene EIF4G2 after vehicle or 100 nM SAG treatment in control and 4H cells. Quantification of GLI1 band intensity shows that, after correction for EIF4G2 band intensity, the GLI1 expression is increased in 4H neurons (C). Veh = vehicle, AU = arbitrary units. Supplementary Table 4-6 are available at Brain online.
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