Figure 5. Notch-1 inhibition leads to upregulation of nicotinic α4- and α6-receptor subunits in human Müller glia with stem cell characteristics (hMGSCs). A: Exemplary images show mRNA expression of nicotinic acetylcholine receptors (nAChR) α1–4, 6, and 7 in hMGSCs under control conditions, after differentiation with basement membrane protein (BMP) and basic fibroblast growth factor-2 (FGF2) alone or after addition of N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycin t-butyl ester (DAPT; left panel). Following differentiation of hMGSCs with the Notch-1 inhibitors BMP, FGF2, and DAPT (red bars), mRNA expression of the α4 and α6 nicotinic acetylcholine receptors (nAChR) subunits was significantly upregulated in comparison to untreated cells (control, black bars) or after treatment with BMP and FGF2 alone (blue bars), as shown by reverse transcription PCR (RT-PCR; *p<0.05, n=6, right panel). B: Immunohistochemical staining showed an increase in protein expression of the α4 and α6 nicotinic AChR subunits (green) with concomitant upregulation of THY1 by hMGSCs treated with BMP, FGF2, and DAPT, in comparison to control (40× magnification).