1. Date of download: 5/30/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Biotin-Responsive Basal Ganglia Disease in Ethnic Europeans With Novel SLC19A3 Mutations Arch Neurol. 2010;67(1):126-130. doi:10.1001/archneurol.2009.293 Brain magnetic resonance image of patient 1. Axial fluid-attenuated inversion recovery sequences before (A-C), during (D-F), and 7 days after biotin treatment (G-I) of a subacute encephalopathy. In addition to persistent lesions of the caudate nuclei and putamen (B, E, H, arrows), abnormally high signals of mesencephalon (D, arrowhead), cortical-subcortical areas (F, arrowheads) were observed during encephalopathy and disappeared after treatment with biotin (G-I). Figure Legend:

2. Date of download: 5/30/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Biotin-Responsive Basal Ganglia Disease in Ethnic Europeans With Novel SLC19A3 Mutations Arch Neurol. 2010;67(1):126-130. doi:10.1001/archneurol.2009.293 Brain magnetic resonance image of patient 2. Axial fluid-attenuated inversion recovery sequences before (A-C), during (D-F), and 1 month after (G-I) treatment of a subacute encephalopathy. In addition to persistent lesions of the caudate nucleus and putamen (B, E, H, arrows), abnormal cortical high signals (D-F, arrowheads or arrows) were observed during encephalopathy and disappeared after treatment with biotin and thiamine (G-I). Figure Legend:

3. Date of download: 5/30/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Biotin-Responsive Basal Ganglia Disease in Ethnic Europeans With Novel SLC19A3 Mutations Arch Neurol. 2010;67(1):126-130. doi:10.1001/archneurol.2009.293 Sequencing of SLC19A3 in patients 1 and 2 identified 3 novel variants present in the heterozygote state in both patients. A, Detection of 3 heterozygous variants of SLC19A3 in patient 1, also present in patient 2 (not shown). The c.980-38dupA was found in 22 of 94 control individuals in contrast to the 2 other remaining variants. The mutation nomenclature is based on the SLC19A3 complementary DNA reference sequence (rs NM_025243) with +1 corresponding to the A of the ATG translation initiation codon. B, Pedigree of the family and segregation analysis of the 3 SLC19A3 variants. C, Reverse/transcriptase–polymerase chain reaction (RT-PCR) products from the RNA of fibroblasts showing aberrant splicing that results from the c.980-14 A>G mutation. This analysis shows the presence of the normal transcript (325 base pairs [bp]) in samples from a control subject and patients 1 and 2. An aberrant splice variant (132 bp) is found only in patients 1 and 2. Treatment of patients' fibroblasts with emetine stabilizes the 132- bp transcript, which indicates that it is normally degraded by nonsense- mediated decay. Exons are represented by boxes. Exon 4 is skipped in the 132bp transcript. M indicates molecular weight; +, with emetine; −, without emetine; C, control; #1, patient 1; and #2, patient 2. D, The identity of the aberrant RT-PCR–product band was confirmed by DNA sequencing: the c.980-14 A>G mutation causes skipping of exon 4. Figure Legend: