Figure 7.

Astrocyte activation in WT3 derived retinal organoids after drug treatment. (A): Astrocytes (GFAP; red) were observed in all retinal organoids across all conditions. While astrocytes were found underneath the neuroepithelium in the center of retinal organoids in control and ketorolac condition they sprouted out into the neuroepithelium after exposure to digoxin, thioridazine, sildenafil, ethanol, and methanol, suggesting activation of astrocytes. Nuclei were counterstained with Hoechst (blue). Scale bars, 20 μm. (B): A higher percentage of astrocytes was found in retinal organoids treated with digoxin, thioridazine, and dildenafil. Data are shown as an individual percentage for each cell type and condition. The mean (black line) ± SEM (dashed line) of control and ketorolac conditions are shown. (C): CCL2 gene expression (shown on the original UMAP) revealed an increase in the percentage of expressing cells upon thioridazine treatment. (D): An increase in the fraction of cells expressing α-crystallin B (CRYAB; green) expression was noted after digoxin, thioridazine, and sildenafil exposure. Nuclei were counterstained with Hoechst (blue). Scale bars, 20 μm. Hoe, Hoechst.