Extended Data Figure 2: FACS isolation of retinal cell populations. | Nature

Extended Data Figure 2: FACS isolation of retinal cell populations.

From: Rb suppresses human cone-precursor-derived retinoblastoma tumours

Extended Data Figure 2

Retinal cells were isolated according to size, CD133 and CD44 staining. In study 1, cell type compositions in each fraction (a) were determined by immunostaining with cone arrestin and CRX (b, e), NRL (c), and nestin and PAX6 (d, f). In study 2, cell type compositions (i) were determined by immunostaining with RXRγ and CRX, nestin and CHX10, nestin and PAX6, and CRALBP (j, k). The percentages of the predominant cell types in each population (a, i) and marker specificities (g) are indicated. h, Cone-specific co-staining of cone arrestin and GNAT2 (top) and cone-specific co-staining of RXRγ and CRX (bottom) in FW19 retina. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer. l, Co-staining of cells for EdU with cone arrestin and CRX or with RXRγ and CRX 14 days after transduction of the cone-enriched medium plus large CD133hi CD44 population isolated as in ik with two RB1 shRNAs (yellow arrows) but not with the scrambled control (white arrows). In both studies, CD133hi CD44 medium and large size populations mainly consisted of cells expressing cone markers (CRX and cone arrestin, or CRX and RXRγ). The CD133hi CD44 small population mainly consisted of cells expressing a rod marker (NRL) with a variable proportion expressing cone markers. All CD133lo CD44+ populations mainly consisted of cells co-expressing RPC and glial markers (nestin and PAX6, nestin and CHX10, or CRALBP). The CD133lo CD44 small size population consisted of cells with diverse immunophenotypes. Values and error bars are mean and s.d. of triplicate assays. Scale bars, 30 μm.

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