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Additional Supporting Information may be found in the online version of this article.

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sc-12-0163_sm_SupplFigure1.tif264KSupplemental Figure 1. Calcium imaging of rods at different developmental stages in slice culture preparations. (a) Postnatal day (P)5, (b) P8, and (c) adult retinas were sliced at 150 μm thickness. Retinal slices were stained with Fura-2 and stimulated with 80 mM KCl. (d) Increases in [Ca2+]i (nM) were calculated from each baseline. Data represent mean ± S.D. *p < 0.05.
sc-12-0163_sm_SupplFigure2.tif727KSupplemental Figure 2. Membrane current recording in rods at different developmental stages in slice culture preparations. Membrane current in Nrlp-eGFP positive cells was recorded at postnatal day (P)5 or P8 (a). Recorded cells were identified by Alexa568 in the electrode solution. (c-f) Membrane current of Nrlp-eGFP-positive cells in (c) P5 and (d) P8 retina were recorded by membrane voltage-clamp from -100 mV to 20 mV, with 10 mV increments. From (c) and (d), I-V curves were plotted in (e) and (f), respectively. (b) Hyperpolarization-activated current (Ih) at -100 mV and outward current at 10 mV in developing rods were compared with those in adult rods. Data represent mean ± S.D. *p < 0.05.
sc-12-0163_sm_SupplFigure3.tif2383KSupplemental Figure 3. Calcium imaging of grafted rods in Crx-KO mouse retina. (a) Nrlp-eGFP positive cell in flat-mounted Crx-KO mouse retina. Arrows indicate Nrlp-eGFP positive cell clusters. (b, c) Fura-2 ratio imaging of grafted cells in flat mounted Crx-KO mouse retina at 0 sec (b) and 120 sec (c). Red color indicates high concentration of intracellular Ca2+. (d) Nrlp-eGFP positive graft cells showed Ca2+ responses to high-K+ stimulation.
sc-12-0163_sm_SupplFigure4.tif1449KSupplemental Figure 4. Mouse iPS cells derived from Nrlp-eGFP mice differentiate into all three germ layers in the immunodeficient mice. (a) Colonies of Nrlp-eGFP mouse iPS cells. (b-d) Nrlp-eGFP mouse iPS cells were injected into the testis of immunodeficient (SCID) mice, and produced various types of tissues derived from three germ layers. Scale bars, 200 μm in (a), 100 μm in (b), (c), and (d).
sc-12-0163_sm_SupplFigure5.tif792KSupplemental Figure 5. Mouse iPS cell-derived Nrlp-eGFP-positive cells show hyperpolarization–activated current. Membrane current in (a) control or (b) ZD7288-treated iPS cell-derived Nrlp-eGFP-positive cells were recorded by membrane voltage-clamp from -100 mV (red line) to 10 mV, with 10 mV increments. (c) I-V curve was plotted from 4 trials. The voltage-dependent inward current recorded at membrane hyperpolarization was suppressed by treatment with ZD7288, an HCN channel blocker. Data represent mean ± S.D. *p < 0.05.
sc-12-0163_sm_SupplSvideo1.avi46086KSupplemental Online Video 1. Fura-2 Ca2+ imaging of retinal cells in retinal slice preparation. Sliced retinas were stained with Fura-2. 100 μM glutamate (at 0:48 min) and 80 mM KCl (at 2:13 min) were added to the medium. Interval = 5.0 sec, 20 frames/sec, total time = 3.7 min (45 frames).
sc-12-0163_sm_SupplSvideo2.avi51206KSupplemental Online Video 2. Fura-2 Ca2+ imaging of Nrlp-eGFP-positive developing rods. Retinal cells from postnatal day (P)3 mice were cultured on glass-base dishes for 2-5 days. Retinal cells were stained with Fura-2. 100 μM glutamate (at 1:00 min) and 80 mM KCl (at 5:40 min) were added to the medium. Interval = 12.0 sec, 20 frames/sec, total time = 9.7 min (50 frames).
sc-12-0163_sm_SupplSvideo3.avi115443KSupplemental Online Video 3. Fura-2 Ca2+ imaging of donor cells in retinal slice preparation. Region of interest (ROI) #1-3 in the video correspond to the same ROI in Figure 3C and indicate donor cells in the wild type retina. 80 mM KCl (at 0:30 min) was added to the medium. Interval = 4.7 sec, 20 frames/sec, total time = 2.0 min (37 frames).
sc-12-0163_sm_SupplTable1.pdf83KSupplementary Table 1.

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