Copyright © 2010 American Medical Association. All rights reserved.

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OCT= Optical Coherence Tomography
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Copyright © 2010 American Medical Association. All rights reserved. From: Leber Hereditary Optic Neuropathy Gene Therapy Clinical Trial RecruitmentYear 1 Arch Ophthalmol. 2010;128(9):1129-1135. doi:10.1001/archophthalmol.2010.201 Figure Legend: Illustration of allotopic strategy. Adeno-associated viruses (AAVs) deliver a nuclear-encoded version of ND4 to the nucleus of the cell (step 1). Messenger RNA (mRNA) is transcribed in the nucleus (step 2). The mRNA diffuses into the cytoplasm (step 3), where it directs the synthesis of a mitochondrial-targeting sequence (MTS) attached to the nuclear ND4 protein (nl ND4) that also has a FLAG epitope for immunodetection. The entire fusion protein is translated on cytoplasmic ribosomes (step 4). The MTS directs the MTS/ND4/FLAG fusion protein into the mitochondria where the MTS is cleaved off (step 5). Within the inner mitochondrial matrix the mature protein (ND4/FLAG), without the MTS, is assembled into complex I (step 6). mtDNA indicates mitochondrial DNA; WT, wild type. Date of download: 12/19/2017 Copyright © 2010 American Medical Association. All rights reserved.

Copyright © 2010 American Medical Association. All rights reserved. From: Leber Hereditary Optic Neuropathy Gene Therapy Clinical Trial RecruitmentYear 1 Arch Ophthalmol. 2010;128(9):1129-1135. doi:10.1001/archophthalmol.2010.201 Figure Legend: Alignment of ND4 protein amino acid changes induced by Leber hereditary optic neuropathy mutations. At amino acid 340, arginine (R) is replaced with histidine (H). A secondary mutation at amino acid 320 replaces a glycine (G) with glutamate (E). The top line shows the sequence of the normal ND4 protein. Date of download: 12/19/2017 Copyright © 2010 American Medical Association. All rights reserved.

Copyright © 2010 American Medical Association. All rights reserved. From: Leber Hereditary Optic Neuropathy Gene Therapy Clinical Trial RecruitmentYear 1 Arch Ophthalmol. 2010;128(9):1129-1135. doi:10.1001/archophthalmol.2010.201 Figure Legend: Plots of visual acuity and visual field mean defects. A, Plots of the mean visual acuity in subjects with Leber hereditary optic neuropathy and maternally related family members. B and C, Mean visual acuity of the right eyes (B) and the left eyes (C). D, Plots of Humphrey visual field mean defects. Date of download: 12/19/2017 Copyright © 2010 American Medical Association. All rights reserved.

Copyright © 2010 American Medical Association. All rights reserved. From: Leber Hereditary Optic Neuropathy Gene Therapy Clinical Trial RecruitmentYear 1 Arch Ophthalmol. 2010;128(9):1129-1135. doi:10.1001/archophthalmol.2010.201 Figure Legend: Plots of optical coherence tomography. A, Scatterplot of the Stratus OCT3 (Carl Zeiss Meditec, Dublin, California) average thickness of the retinal nerve fiber layer (RNFL) relative to the months after initial visual loss. All optical coherence tomography was done during the initial visit. B, A plot of the OCT3 average RNFL thickness of affected subjects and carriers. Date of download: 12/19/2017 Copyright © 2010 American Medical Association. All rights reserved.

Copyright © 2010 American Medical Association. All rights reserved. From: Leber Hereditary Optic Neuropathy Gene Therapy Clinical Trial RecruitmentYear 1 Arch Ophthalmol. 2010;128(9):1129-1135. doi:10.1001/archophthalmol.2010.201 Figure Legend: Plots of pattern electroretinogram (PERG) amplitude. A, The PERG amplitude relative to normal is shown for affected subjects and carriers. B and C, The PERG amplitude percentage of normal is shown relative to visual acuity with scatterplots in affected subjects (B) and carriers (C). D and E, The PERG amplitude relative to normal is shown relative to the mean defect on Humphrey visual field tests with scatterplots in affected subjects (D) and carriers (E). F and G, The PERG amplitude relative to normal is shown with scatterplots relative to the Stratus OCT3 (Carl Zeiss Meditec, Dublin, California) retinal nerve fiber layer (RNFL) thickness in affected subjects (F) and carriers (G). AA320 F indicates female with primary G11778A mutation plus secondary mutation in ND4 mitochondrial DNA (mtDNA); AA320 M, male with primary G11778A mutation plus secondary mutation in ND4 mtDNA; No AA320 M, male with primary G11778A mtDNA mutation but no secondary mtDNA mutation; No AA320 F, female with primary G11778A mutation but no secondary mtDNA mutation. Date of download: 12/19/2017 Copyright © 2010 American Medical Association. All rights reserved.