Figure 4.

Altered expression of rod photoreceptor cell markers in LCA7 retinal organoids at D180

(A–Y) Immunofluorescence staining using antibodies against OTX2 (white; A–F′), SAG (green; G–L′), NRL (red; M–R′), and NR2E3 (red; S–X′) are shown for control (CRX^WT; A–B′, G–H′, M–N′, S–T′), CRX^T155ins4/+ (C–D′, I–J′, O–P′, U–V′), and CRX^K88Q/+ (E–F′, K–L′, Q–R′, W–X′) retinal organoids at D180 (n = 9 total organoids from three experimental replicates per line). Nuclei are counterstained with DAPI (blue). OPL, outer plexiform layer; INL/GCL, inner nuclear layer/ganglion cell layer. qRT-PCR data (Y) are shown for CRX^T155ins4/+ (green line) and CRX^K88Q/+ (blue line) at D75, D90, D120, D150, and D180 as fold change compared with control organoids for OTX2, NRL, and NR2E3 (n = 14 total organoids from two experimental replicates per line). The dotted line represents no change compared with control (y = 1). All statistical analyses were performed using one-way ANOVA with a Dunnett test to correct for multiple comparisons in GraphPad Prism 8 software. Scale bar (X and X′), 100 μm. ^∗p < 0.05, ^∗∗∗p < 0.005, ^∗∗∗∗p < 0.001.

See also Figures S3, S4, and S7.