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. 2022 Mar 18;13:837756. doi: 10.3389/fphar.2022.837756

FIGURE 5.

FIGURE 5

Validation of candidate compounds in a chemically induced Gaucher disease model. (A) High throughput imaging of DA neurons with the InCell 6,000 platform for the positive control, CBE, and the candidate compounds. The bottom left image shows the DA neuron isolation process in the custom Cellprofiler pipeline used for image analysis. (B) Hit validation of candidate compounds with 48 h treatment of 500 μM CBE. At 5 dpf, larvae were treated with 0.2% DMSO (positive control), 500 μM CBE (negative control), and the CBE+ 10 μM candidate compounds for 48 h. At 7 dpf, the larvae were imaged with a confocal microscope. The 500 μM CBE showed significant reduction in DA neurons compared to the 0.2% DMSO control (N = 12; p = 0.0012, unpaired t-test). Nepafenac, olmesartan, and aloperine showed significant neuroprotection when co-treated with CBE (N = 10 to 12; one-way ANOVA F = 6.205, p < 0.001, post-hoc Fishers LSD **p < 0.01, ***p < 0.001, unpaired t-test). CBE: Conduritol B epoxide.