Supplementary on-line figure legends Figure S1. Haplotype diversity for 19 SNPs within 137kb upstream of LRRTM1. Diversity within parents from the RD family sample is shown. The SNPs are (from left to right): Lrrprom1, rs1446109, rs6547303, rs1006154, rs1006155, rs1007371, rs920818, rs310784, rs1373264, rs718467, rs1947033, rs728335, rs149248, rs723524, rs1025947, rs985507, rs1444539, rs1444542, rs6747004 (see locations in Table S1). (Lrrprom1 is within the predicted promoter region of LRRTM1 according to the program FirstEF http://rulai.cshl.org/tools/FirstEF/. Best estimate haplotypes were derived using PHAMILY Ackerman et al.: http://archimedes.well.ox.ac.uk/pise/phamily-simple.html . The three SNPs that led to the initial identification of paternal-specific association with relative hand skill are highlighted by asterisks at the bottom. The family of 2-2-2 haplotypes defined by these three SNPs is highlighted by a dashed red surround.
Figure S2. Expression of bi-allelic control genes from human chromosome 2 in A9 hybrid cells, regardless of parental origin. Expression is assessed by RT-PCR. Cell lines with blue identities contain a paternally derived human chromosome 2, those with red identities contain a maternally derived human chromosome 2.
Figure S3. In situ hybridization analysis of Lrrtm1 mRNA expression in E12.5 mouse embryos (A-D) and in adult mouse brain (E-H). Shown are dark-field emulsion autoradiographs and corresponding bright-field images of the sections (counterstained with hematoxylin). The white signal in the dark-field images indicates Lrrtm1 expression. (A-D) In E12.5 mouse, Lrrtm1 is predominantly expressed by postmitotic neurons of the CNS as well as by some non-neuronal tissues (e.g. mesenchyme in head region and around vertebrae). Sections A and C correspond to Plates 29a and 29b, respectively, in the Atlas of Mouse Development (Kaufman MH, 2nd Ed. Academic Press, 1995). (E-H) In adult mouse brain, Lrrtm1 expression is abundant in various thalamic nuclei, in the hippocampus, as well as in the piriform and retrosplenial cortex. Sections E and G correspond to Figs. 35 and 55, respectively, in the Mouse Brain in Stereotaxic Coordinates (Paxinos G & Franklin K, 2nd Ed. Academic Press, 2001). Abbreviations: 3V, third ventricle; 4V fourth ventricle; Cg, (anterior) cingulate cortex; CPu, caudate-putamen; Ent, entorhinal cortex; GE, ganglionic eminence, HC, hippocampus; LV, lateral ventricle; Me, medulla; PAG, periaqueductal gray; Pir, piriform cortex; Po, pons; RSG, retrosplenial granular cortex; Th, thalamus; SC, spinal cord; SuC, superior colliculus; VTA, ventral tegmental area. Scale bar = 500 µm.
Figure S4. Northern blot analysis of LRRTM1 mRNA expression in adult human brain regions using BD Biosciences Multiple Tissue Northern (MTN) blots II (left) and V (right). A band of approximately 3 kb corresponding to LRRTM1 transcript is detected (top). Control hybridization with -Actin probe is shown (bottom).
Figure S5. Biallelic expression of Lrrtm1 mRNA in F1 mice between B6 and JF strains. gDNA represents genomic DNA.
Figure S6. LRRTM1 localizes in the ER in mammalian cells. MRC-5 SV2 cells were transfected with plasmids pcDmL1 (top) or pcDmL1m (bottom) and processed for IF. Both untagged and C-terminal myc+His tagged LRRTM1 proteins (green), detected with specific anti-peptide 126, co-localize with the ER marker BAP31 (red). Similar results were obtained also with 293T cells (data not shown).