Figure 2. Expression of retinal ganglion cell (RGC) markers following Notch-1 inhibition in human Müller glia with stem cell characteristics
(hMGSCs) is consistent with the acquisition of a neural phenotype. A: Inhibition of Notch-1 by treatment with basement membrane protein (BMP), basic fibroblast growth factor-2 (FGF2) and N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycin
t-butyl ester (DAPT) attenuated mRNA expression of HES1 (***p<0.001, n=4), while significantly upregulating BRN3b mRNA, a marker of RGC precursors (**p<0.01, n=4), as shown by quantitative and conventional reverse transcription PCR (RT-PCR),
respectively. B: In untreated hMGSCs, the Müller cell markers vimentin and CRALBP (green) were highly expressed, while THY1, βIII-tubulin,
and ISL-1 (red), which are characteristic of RGCs, were undetectable or found at low levels (upper panel). After differentiation
of hMGSCs by Notch-1 inhibition, the expression of vimentin and CRALBP (green) was attenuated, while that of THY1, βIII-tubulin,
and ISL-1 was augmented (red, lower panel).