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. 2019 Sep 19;13(4):747–760. doi: 10.1016/j.stemcr.2019.08.012

Figure 3.

Figure 3

Characterization of CCND1 Function in RPC Commitment

(A) Immunostaining of CCND1, Ki67, and ASCL1 in the central and peripheral day-35 retinal organoids. Scale bars, 25 μm.

(B) Schematic presentation of CCND1 inducible overexpression experiment. Retinal organoids were transduced by lentiviral particles carrying a Tet-On CCND1 overexpression system that can overexpress ectopic CCND1 upon the addition of doxycycline (Dox). We specifically overexpress CCND1 in retinal organoids from day 28 to day 32 by adding Dox at day 28 and removing Dox after day 32.

(C) Expression patterns of ASCL1, ISLET1, NEUROD1, and BLIMP1 in CCND1-overexpressing (CCND1 OE) retinal organoids and control organoids at day 32. Scale bars, 50 μm.

(D) Quantification of Marker+ cells in CCND1 OE retinal organoids and control organoids (mean ± SD; n ≥ 5 organoids from two differentiations) at day 32. ∗p < 0.05, ∗∗p < 0.001 (unpaired t test).

See also Figure S3.