Figure 5.

qRT-PCR and immunofluorescence staining reveal significant downregulation of late photoreceptor cell markers in LCA7 organoids at D180

(A–S′) qRT-PCR data (A) are shown for CRX^WT (gray lines), CRX^T155ins4/+ (light gray), and CRX^K88Q/+ (dark gray) at D180 as fold change compared with control organoids for RHO, OPN1SW, OPN1MW, and OPN1LW (n = 14 total organoids from two experimental replicates per line). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.005, ^∗∗∗∗p < 0.001. Immunofluorescence staining using antibodies against RHO (green; B–G′), S-opsin (red; H–M′), and M/L-opsin (green; N–S′) is shown for control (CRX^WT; B–C′, H–I′, N–O′), CRX^T155ins4/+ (D–E′, J–K′, P–Q′), and CRX^K88Q/+ (F–G′, L–M′, R–S′) retinal organoids at D180 (n = 9 total organoids from three experimental replicates per line). Nuclei are counterstained with DAPI (blue). Scale bar (S and S′), 100 μm. OPL, outer plexiform layer; INL/GCL, inner nuclear layer/ganglion cell layer. All statistical analyses were performed using one-way ANOVA with a Dunnett test to correct for multiple comparisons in GraphPad Prism 8 software.

See also Figures S3 and S4.