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. 2019 Aug 7;13:361. doi: 10.3389/fncel.2019.00361

FIGURE 4.

FIGURE 4

USH2A mutation iPSCs generate retinal organoids with defective retinal neuroepithelial and RPC development. (A) The thickness of retinal neuroepithelium in RP-organoids at days 15–22. (B) Brightfield images of developing retinal neuroepithelium at day 18. (C) Immunofluorescence staining shows the cells in retinal organoids co-expressing RAX and CDH2 (N-cadherin) immunoreactivity at day 10. (D) Immunofluorescence images shows the cells in NR organoids expressing CHX10 immunoreactivity at day 18. (E,F) Immunostaining of NR organoids showing the expression of specific markers PAX6, N-cadherin, and aPKC at day 18. (G) qPCR analysis reveals mRNA levels of RPCs specific markers PAX6, RAX, SIX6, and CHX10 (GAPDH gene as a control). (H,I) Neural retina development and RPCs related DEGs at day 34 were normalized by Z-scores. Red represents upregulated expression. Blue represents downregulated expression, Bar is represents Z-score. (n = 3 independent experiments; each experiment need 2–4 organoids; scale bar = 200 μm).