PMC:1084331 / 47038-51048 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/1084331","sourcedb":"PMC","sourceid":"1084331","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1084331","text":"Supporting Information\nFigure S1 Gene Expression Analysis in DRG Neurons of Er81EWS-Pea3 Mice\nAnalysis of TrkC expression by in situ hybridization (A and E), and Runx3 (red; B and F), Brn3A (red; C and G), Isl1 (red; D and H), p75 (red; I and M), TrkA (red; J and N), CGRP (red; K and O), Calretinin (CR; red; L and P), and LacZ (green; B–D and F–P) by immunohistochemistry on E16.5 lumbar DRG of Er81NLZ/+ (A–D and I–L) and Er81EWS-Pea3/− (E–H and M–P) embryos.\nScale bar: (A, B, E, F, L, and P), 70 μm; (C, D, G, H, I–K, and M–O), 75 μm.\n(4.9 MB CDR).\nClick here for additional data file.\nFigure S2 Ia Proprioceptive Afferents Make Functional Connections with Motor Neurons in Er81EWS-Pea3 Mutants\n(A and B) Representative traces from intracellular recordings measuring Ia afferent monosynaptic input to quadriceps motor neurons evoked by suprathreshold stimulation of the quadriceps nerve in wild-type (A) and Er81EWS-Pea3/− mutant (B) animals.\n(C) Average monosynaptic amplitudes (± standard error of the mean) from all recorded cells (wild-type, n = 11; mutant, n = 8).\n(20 KB CDR).\nClick here for additional data file.\nFigure S3 Generation of Mice Expressing EWS-Pea3 or mGFP in Early Post-Mitotic DRG Neurons\n(A) Top panel shows organization of the Tau genomic locus in the region targeted by homologous recombination in analogy to Tucker et al. [41]. Exons 1–3 are shown as light blue boxes, and the Tau start codon in exon 2 is indicated as ATG. The probe used to detect homologous recombination is shown as a grey box. Middle and bottom panels show Tau locus after homologous recombination to integrate targeting cassettes (green) into exon 2 with coincident elimination the endogenous Tau start codon. The integrated targeting cassettes allow for conditional expression of EWS-Pea3 and NLS-LacZ (NLZ) (middle) or mGFP and NLS-LacZ (bottom) upon Cre-recombinase-mediated activation. In the absence of Cre recombinase, a transcriptional stop sequence flanked by loxP sites inhibits expression of the respective transgenes from their start codons (ATG in grey).\n(B) Southern blot analysis of TauEWS-Pea3/+ and TaumGFP/+ genomic DNA to detect the mutant allele.\n(C) In the presence of Cre recombinase, the transcriptional stop sequence in the cassettes integrated into the Tau locus is removed. Expression of EWS-Pea3 and NLS-LacZ (top) or mGFP and NLS-LacZ (bottom) can now occur in neurons coincidently expressing Cre recombinase and Tau (indicated as ATG in green).\n(D–L) Expression of Isl1 (D, G, and J), EWS-Pea3 (E and H), GFP (K), or LacZ (F, I, and L), in E12 (D–I) or E13.5 (J–L) DRG neurons of wild-type (D–F), TauEWS-Pea3/+ Isl1Cre/+ (G–I), and TaumGFP/+ Isl1Cre/+ (J–L) embryos.\nScale bar: (D–F), 40 μm; (G–I), 35 μm; (J–K), 50 μm.\n(1.5 MB CDR).\nClick here for additional data file.\nFigure S4 Generation of PVCre Mice\n(A) Above is the organization of the PV genomic locus. Exons are schematically illustrated as light blue boxes, where exon 2 contains the start codon (ATG) and exon 5 contains the stop codon (STOP). Probe to screen for homologous recombination is shown as grey box. Below is a schematic diagram to show the PV locus after the integration of an IRES-Cre cassette (green) 3′ to the translational stop codon of PV using homologous recombination in ES cells.\n(B) Southern blot analysis of PVCre wild-type (+/+) and heterozygous (+/−) genomic DNA using the probe indicated in (A).\n(C and D) Expression of GFP (green) and LacZ (red; C) or PV (red; D) in P0 TaumGFP/+ PVCre/+ mice. Note that more than 90% of PV+ neurons coexpress GFP (D; data not shown).\nScale bar: (C and D), 50 μm.\n(2 MB CDR).\nClick here for additional data file.\nFigure S5 Gene Expression Analysis upon Precocious Induction of EWS-Pea3 in DRG Neurons\nImmunohistochemical analysis of PV (A and D), Calretinin (B and E), and Calbindin (C and F) expression on E16.5 lumbar DRG of wild-type (A–C) and TauEWS-Pea3/+ Isl1Cre/+ (D–F) embryos.\nScale bar: 80 μm.\n(950 KB CDR).\nClick here for additional data file.","divisions":[{"label":"title","span":{"begin":0,"end":22}},{"label":"label","span":{"begin":23,"end":32}},{"label":"caption","span":{"begin":34,"end":554}},{"label":"title","span":{"begin":34,"end":94}},{"label":"p","span":{"begin":95,"end":463}},{"label":"p","span":{"begin":464,"end":540}},{"label":"p","span":{"begin":541,"end":554}},{"label":"caption","span":{"begin":555,"end":591}},{"label":"p","span":{"begin":555,"end":591}},{"label":"label","span":{"begin":592,"end":601}},{"label":"caption","span":{"begin":603,"end":1089}},{"label":"title","span":{"begin":603,"end":701}},{"label":"p","span":{"begin":702,"end":949}},{"label":"p","span":{"begin":950,"end":1076}},{"label":"p","span":{"begin":1077,"end":1089}},{"label":"caption","span":{"begin":1090,"end":1126}},{"label":"p","span":{"begin":1090,"end":1126}},{"label":"label","span":{"begin":1127,"end":1136}},{"label":"caption","span":{"begin":1138,"end":2767}},{"label":"title","span":{"begin":1138,"end":1218}},{"label":"p","span":{"begin":1219,"end":2072}},{"label":"p","span":{"begin":2073,"end":2171}},{"label":"p","span":{"begin":2172,"end":2478}},{"label":"p","span":{"begin":2479,"end":2700}},{"label":"p","span":{"begin":2701,"end":2753}},{"label":"p","span":{"begin":2754,"end":2767}},{"label":"caption","span":{"begin":2768,"end":2804}},{"label":"p","span":{"begin":2768,"end":2804}},{"label":"label","span":{"begin":2805,"end":2814}},{"label":"caption","span":{"begin":2816,"end":3630}},{"label":"title","span":{"begin":2816,"end":2840}},{"label":"p","span":{"begin":2841,"end":3295}},{"label":"p","span":{"begin":3296,"end":3416}},{"label":"p","span":{"begin":3417,"end":3589}},{"label":"p","span":{"begin":3590,"end":3618}},{"label":"p","span":{"begin":3619,"end":3630}},{"label":"caption","span":{"begin":3631,"end":3667}},{"label":"p","span":{"begin":3631,"end":3667}},{"label":"label","span":{"begin":3668,"end":3677}},{"label":"caption","span":{"begin":3679,"end":3973}},{"label":"title","span":{"begin":3679,"end":3756}},{"label":"p","span":{"begin":3757,"end":3941}},{"label":"p","span":{"begin":3942,"end":3959}},{"label":"p","span":{"begin":3960,"end":3973}}],"tracks":[{"project":"2_test","denotations":[{"id":"15836427-11135642-84719690","span":{"begin":1357,"end":1359},"obj":"11135642"},{"id":"T56896","span":{"begin":1357,"end":1359},"obj":"11135642"}],"attributes":[{"subj":"15836427-11135642-84719690","pred":"source","obj":"2_test"},{"subj":"T56896","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#ec93d2","default":true}]}]}}