Fig 2: iPSC-MSCs administration prevented visual function decline and loss of retinal ganglion cell. A & B. After intravitreal administration, retinal function analysis by electroretinogram (ERG). (A) 1 week and (B) 4 weeks post injection, (i) a-wave, (ii) b-wave and (iii) PhRN amplitudes was recorded. (n=10). C. Density of RGC in the RGC layer immune labeled with TUBB3 antibody in WT mice and KO mice treated with PBS, MSC and MSC(R) were compared both at one week (upper lane) and four weeks (lower lane) after intravitreal injection. (n=5). Data information: In (A-C), data are presented as mean ± SD. *p<0.5, **p<0.01, ***p<0.001, ns, not significant (One-way ANOVA,).

Fig 3: Detection of human mitochondrial within Ndufs4 deficient mice. A. PCR results of retina tissue at 96 hours, 1 week and 4 weeks post treatment. DNA from human iPSC-MSCs, mice tissue and mixture (human cells + mice tissue) served as control to validate the primer amplification efficiency in panel (Ai). In all the time points, except H-nDNA, the other three including H-mDNA, M-nDNA, and M-mDNA could be detected in the mouse retinal samples (Aii). H-nDNA: human specific nuclear DNA, H-mDNA: human specific mitochondrial DNA, M-nDNA: mice specific nuclear DNA, M-mDNA: mice specific mitochondrial DNA. To confirm, HNA signal was detected in the VRI only but not GCL in the flat mounted mouse retinas at 1-week post injection (Aiii). Scale bar: 50 µm. B. At 96 hours post injection, positive mito-GFP but negative TUBB3 expression of iPSC-MSCs was detected in VRI (Green; arrows, Bi). These mito-GFP signals in VRI were overlapped with anti-hMito immunostained mitochondria (Red; arrows,Bii). Arrow head indicates human mitochondria labelled with anti-hMito but without mito-GFP in iPSC-MSC (Bii). In GCL the transferred mitochondria expressingmito-GFP signals (arrow heads) in the TUBB3 positive cells (Biii). Anti-hMito signals were detected in the cytoplasm of the Brn3a positive RGC cells (Red; arrow heads; Biv). Scale bars: 10 µm. C. Z-stack images showed iPSC-MSCs on VRI at different time post injection. iPSC-MSC cells expressing LV-Mito-GFP signals and immunostained anti-human mitochondria (anti-hMito, red) signals were detected at 96 hours and 1 week post injection but not 4 weeks. Anti-hMito signals existed in GCL surrounding RGCs were detected up to 4 weeks in the X-Z panel; arrow heads. DAPI (Blue). HNA - human nuclear antigen; VRI - vitreoretinal interface. Scale bars: 50 µm.