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  • 88
    Zyagen human brain complementary dna cdna
    Human Brain Complementary Dna Cdna, supplied by Zyagen, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    BioChain Institute human brain complementary dna
    Human Brain Complementary Dna, supplied by BioChain Institute, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa human brain cdna
    Analysis of ADAM 23 expression in human tissues and cell lines. (A) Approximately 2 μg of polyadenylated RNA from the indicated tissues were analyzed by hybridization with the full-length <t>cDNA</t> isolated for human ADAM 23. The positions of RNA size markers are shown. Filters were subsequently hybridized with a human actin probe to ascertain the differences in RNA loading among the different samples. (B) Reverse <t>transcription-PCR</t> analysis of ADAM 23 expression in human neuroblastoma NB100 and SH-S y 5 y cells. N1 and N2 indicate negative controls for each cell line. M, molecular size markers.
    Human Brain Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher human fetal brain complementary dna library
    Analysis of ADAM 23 expression in human tissues and cell lines. (A) Approximately 2 μg of polyadenylated RNA from the indicated tissues were analyzed by hybridization with the full-length <t>cDNA</t> isolated for human ADAM 23. The positions of RNA size markers are shown. Filters were subsequently hybridized with a human actin probe to ascertain the differences in RNA loading among the different samples. (B) Reverse <t>transcription-PCR</t> analysis of ADAM 23 expression in human neuroblastoma NB100 and SH-S y 5 y cells. N1 and N2 indicate negative controls for each cell line. M, molecular size markers.
    Human Fetal Brain Complementary Dna Library, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Becton Dickinson human brain cdna
    Analysis of ADAM 23 expression in human tissues and cell lines. (A) Approximately 2 μg of polyadenylated RNA from the indicated tissues were analyzed by hybridization with the full-length <t>cDNA</t> isolated for human ADAM 23. The positions of RNA size markers are shown. Filters were subsequently hybridized with a human actin probe to ascertain the differences in RNA loading among the different samples. (B) Reverse <t>transcription-PCR</t> analysis of ADAM 23 expression in human neuroblastoma NB100 and SH-S y 5 y cells. N1 and N2 indicate negative controls for each cell line. M, molecular size markers.
    Human Brain Cdna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 89/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    OriGene human brain cdna
    Annotated variants of the GRN 5′ <t>UTR.</t> A , sequence of the 219 nucleotides of the GRN 5′ UTR derived from human brain <t>cDNA.</t> This annotated variant ( NM002087 ) and start sites of predicted shorter variants ( XM005257255 and XM005257254 ) are indicated, as well as a predicted splice variant ( XM005257253 ) containing a short sequence stretch of exon 1, followed by an insertion of 271 nucleotides of intron 1 ( green box ) at the exon 1/2 boundary. 5′ ends identified by rapid amplification of cDNA ends are marked by arrows. Green arrows indicate mRNAs containing the intron 1-derived sequence stretch. ORFs are highlighted in red , and AUG start codons are circled. B , schematic of the 219-nucleotide-long GRN 5′ UTR. Two AUG initiation codons (positions 90 and 120), the corresponding uORF, and the stop codon (nucleotides 159–161) are indicated. C , schematic of the alternative spliced GRN 5′ UTR ( XM005257253 ). This splice variant contains a short stretch of exon 1 (nucleotides 190–212), followed by 271 nucleotides derived from intron 1 containing two AUGs ( circled ) and one short uORF ( 156–176 ).
    Human Brain Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher human brain cdna
    Expression Studies of <t>SYT14/Syt14</t> <t>cDNA</t> in Human and Mouse Tissues and Localization of SYT14 in Transfected COS-1 Cells
    Human Brain Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Analysis of ADAM 23 expression in human tissues and cell lines. (A) Approximately 2 μg of polyadenylated RNA from the indicated tissues were analyzed by hybridization with the full-length cDNA isolated for human ADAM 23. The positions of RNA size markers are shown. Filters were subsequently hybridized with a human actin probe to ascertain the differences in RNA loading among the different samples. (B) Reverse transcription-PCR analysis of ADAM 23 expression in human neuroblastoma NB100 and SH-S y 5 y cells. N1 and N2 indicate negative controls for each cell line. M, molecular size markers.

    Journal: Molecular Biology of the Cell

    Article Title: ADAM 23/MDC3, a Human Disintegrin That Promotes Cell Adhesion via Interaction with the ?v?3 Integrin through an RGD-independent Mechanism

    doi:

    Figure Lengend Snippet: Analysis of ADAM 23 expression in human tissues and cell lines. (A) Approximately 2 μg of polyadenylated RNA from the indicated tissues were analyzed by hybridization with the full-length cDNA isolated for human ADAM 23. The positions of RNA size markers are shown. Filters were subsequently hybridized with a human actin probe to ascertain the differences in RNA loading among the different samples. (B) Reverse transcription-PCR analysis of ADAM 23 expression in human neuroblastoma NB100 and SH-S y 5 y cells. N1 and N2 indicate negative controls for each cell line. M, molecular size markers.

    Article Snippet: To obtain this DNA fragment, we performed PCR amplification of a human brain cDNA ( Clontech ) with two specific primers, 5′-CAACAAAGCTATTTGAGCCCACGG and 5′-TTGGTGGGCACTGACCAGAGTCT, derived from the sequence.

    Techniques: Expressing, Hybridization, Isolation, Polymerase Chain Reaction

    Characterization of BPAG1n4. (A) Schematic of the domain structures of BPAG1 isoforms. I, first exon of each isoform; BPAG1e, the epithelial isoform; ABD, actin-binding domain; IF-BD, intermediate filament-binding domain; M1, microtubule-binding domain. Amino acid residue numbers denote functional domain boundaries in BPAG1n4. Individual domains and regions used in this work are illustrated below. Anti-BPAG1n recognizes all BPAG1 isoforms. (B) Northern blot analysis of mouse dorsal root ganglion (DRG) RNA. (probes) A 1.6-kb cDNA head domain fragment (top; 4-d exposure); GAPDH as internal control (bottom; 1-h exposure). (C) Protein expression of BPAG1n4 in mouse brain. Blot was probed with anti-BPiso4. COS-7 (lane 3) and NIH 3T3 cells (lane 4) were transfected with an expression construct for n4-92 (A, bottom) encompassing the epitope for anti-BPiso4. (untrans.) Untransfected COS-7 (lane 5) and NIH cells (lane 6). (D–E) ImmunoEM reveals subcellular localization of BPAG1n4 in dorsal roots. (D) BPAG1n4 labeling with anti-BPiso4 was visualized with 12-nm gold particles conjugated with secondary antibody. White arrows indicate BPAG1n4 on vesicle-like structures in association with microtubules. (E) Negative control, secondary antibodies alone. Bar: (D) 300 nm; (E) 400 nm.

    Journal: The Journal of Cell Biology

    Article Title: BPAG1n4 is essential for retrograde axonal transport in sensory neurons

    doi: 10.1083/jcb.200306075

    Figure Lengend Snippet: Characterization of BPAG1n4. (A) Schematic of the domain structures of BPAG1 isoforms. I, first exon of each isoform; BPAG1e, the epithelial isoform; ABD, actin-binding domain; IF-BD, intermediate filament-binding domain; M1, microtubule-binding domain. Amino acid residue numbers denote functional domain boundaries in BPAG1n4. Individual domains and regions used in this work are illustrated below. Anti-BPAG1n recognizes all BPAG1 isoforms. (B) Northern blot analysis of mouse dorsal root ganglion (DRG) RNA. (probes) A 1.6-kb cDNA head domain fragment (top; 4-d exposure); GAPDH as internal control (bottom; 1-h exposure). (C) Protein expression of BPAG1n4 in mouse brain. Blot was probed with anti-BPiso4. COS-7 (lane 3) and NIH 3T3 cells (lane 4) were transfected with an expression construct for n4-92 (A, bottom) encompassing the epitope for anti-BPiso4. (untrans.) Untransfected COS-7 (lane 5) and NIH cells (lane 6). (D–E) ImmunoEM reveals subcellular localization of BPAG1n4 in dorsal roots. (D) BPAG1n4 labeling with anti-BPiso4 was visualized with 12-nm gold particles conjugated with secondary antibody. White arrows indicate BPAG1n4 on vesicle-like structures in association with microtubules. (E) Negative control, secondary antibodies alone. Bar: (D) 300 nm; (E) 400 nm.

    Article Snippet: Molecular cloning of BPAG1n4 Complementary DNAs of BPAG1n4 were amplified by rapid amplification of cDNA ends–PCR in human brain Marathon-Ready cDNA (CLONTECH Laboratories, Inc.).

    Techniques: Binding Assay, Functional Assay, Northern Blot, Expressing, Transfection, Construct, Labeling, Negative Control

    An SVA retrotransposal insertion induces abnormal splicing in FCMD a, Expression analysis of various regions of fukutin mRNA in lymphoblasts. Gray bar, the ratio of RT-PCR product in FCMD patients relative to the normal control; Numbers on the X axis, nucleotide positions of both forward and reverse primers in fukutin . Error bars, s.e.m. b, Long range PCR using primers flanking the expression-decreasing area (nucleotide position 1061 to 5941) detected a 3-kb PCR product in FCMD lymphoblast cDNA (open arrow) and 8-kb product in FCMD genomic DNA (closed arrow). In the normal control, cDNA and genomic DNA both showed 5-kb PCR products. The 8-kb band was weak probably because VNTR region of SVA is GC-rich (82%). c, Schematic representation of genomic DNA and cDNA in FCMD. Black and white arrows, forward and reverse sequencing primers. The intronic sequence in FCMD is indicated in lower case. The authentic stop codon is colored in red, and the new stop codon is colored in blue. d, e, Northern blot analysis of fukutin in human lymphoblasts ( d ) and model mice ( e ). F, FCMD; N, nomal control. The wild-type mouse fukutin mRNA was detected at a size of 6.1 kb. Both skeletal muscle (left) and brain (right) showed smaller, abnormal bands (open arrows) in Hp/Hp mice. Wt, wild type; Hn, Hn/Hn mice; Hp, Hp/Hp mice. f, Schematic representation of genomic DNA and cDNA in ARH ( LDLRAP1 , left), NLSDM ( PNPLA2 , middle), and human ( AB627340 , right).

    Journal: Nature

    Article Title: Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy

    doi: 10.1038/nature10456

    Figure Lengend Snippet: An SVA retrotransposal insertion induces abnormal splicing in FCMD a, Expression analysis of various regions of fukutin mRNA in lymphoblasts. Gray bar, the ratio of RT-PCR product in FCMD patients relative to the normal control; Numbers on the X axis, nucleotide positions of both forward and reverse primers in fukutin . Error bars, s.e.m. b, Long range PCR using primers flanking the expression-decreasing area (nucleotide position 1061 to 5941) detected a 3-kb PCR product in FCMD lymphoblast cDNA (open arrow) and 8-kb product in FCMD genomic DNA (closed arrow). In the normal control, cDNA and genomic DNA both showed 5-kb PCR products. The 8-kb band was weak probably because VNTR region of SVA is GC-rich (82%). c, Schematic representation of genomic DNA and cDNA in FCMD. Black and white arrows, forward and reverse sequencing primers. The intronic sequence in FCMD is indicated in lower case. The authentic stop codon is colored in red, and the new stop codon is colored in blue. d, e, Northern blot analysis of fukutin in human lymphoblasts ( d ) and model mice ( e ). F, FCMD; N, nomal control. The wild-type mouse fukutin mRNA was detected at a size of 6.1 kb. Both skeletal muscle (left) and brain (right) showed smaller, abnormal bands (open arrows) in Hp/Hp mice. Wt, wild type; Hn, Hn/Hn mice; Hp, Hp/Hp mice. f, Schematic representation of genomic DNA and cDNA in ARH ( LDLRAP1 , left), NLSDM ( PNPLA2 , middle), and human ( AB627340 , right).

    Article Snippet: To detect abnormally-spliced RT-PCR products from FCMD, ARH, and NLSDM patients, and from human brain AB627340 cDNA, long range PCR was performed using LA Taq with LA Taq Buffer II (Takara), adding dimethyl sulfoxide and 7-deaza-dGTP (Roche).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Sequencing, Northern Blot, Mouse Assay

    Expression of TBP-2 in normal and tumor tissues. ( a ) Multiple tissue Northern blots, containing poly(A) + RNA from the indicated tissues (CLONTECH), were hybridized with a 1.1-kb 32 P-labeled TBP-2 cDNA probe ( Upper ). The bolts were rehybridized with a 2.0-kb probe for β-actin, as a control for loading ( Lower ). sk. muscle, skeletal muscle; per. bl. leuk., peripheral blood leukocytes. ( b ) A dot blot containing matched samples of cDNA extracted from normal human tissues and tumors (CLONTECH) was hybridized with a 1.1-kb 32 P-labeled TBP-2 cDNA probe. Samples of colon and breast tumors (T) are shown, with the cDNA from the normal tissue (N) shown directly above each corresponding tumor sample.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin

    doi: 10.1073/pnas.182372299

    Figure Lengend Snippet: Expression of TBP-2 in normal and tumor tissues. ( a ) Multiple tissue Northern blots, containing poly(A) + RNA from the indicated tissues (CLONTECH), were hybridized with a 1.1-kb 32 P-labeled TBP-2 cDNA probe ( Upper ). The bolts were rehybridized with a 2.0-kb probe for β-actin, as a control for loading ( Lower ). sk. muscle, skeletal muscle; per. bl. leuk., peripheral blood leukocytes. ( b ) A dot blot containing matched samples of cDNA extracted from normal human tissues and tumors (CLONTECH) was hybridized with a 1.1-kb 32 P-labeled TBP-2 cDNA probe. Samples of colon and breast tumors (T) are shown, with the cDNA from the normal tissue (N) shown directly above each corresponding tumor sample.

    Article Snippet: Rapid amplification of cDNA ends was performed to determine the transcriptional start site of the TBP-2 gene, using TBP-2-specific primer 1 (5′-GTTGGTTTTAAGAGTTAGAAATGACGG-3′) and nested primer 2 (5′-TAAGGTATTCTTAAGCAGTTTGAGC-3′) with Marathon-ready human brain cDNA (CLONTECH), according to the manufacturer's instructions.

    Techniques: Expressing, Northern Blot, Labeling, Dot Blot

    Expression of TRX in transformed cells cultured with SAHA. T24 bladder carcinoma cells were cultured with vehicle alone (0) or 2.5 or 5 μM SAHA for 6, 15, or 24 h. RNA was extracted and analyzed by Northern blotting for levels of TRX, using a 500-bp 32 P-labeled cDNA probe ( Top ). The blots were subsequently rehybridized with the 1.1-kb 32 P-labeled TBP-2 cDNA probe to confirm induction of TBP-2 ( Middle ) and a γ- 32 P-labeled 18S oligonucleotide probe to indicate RNA loading ( Bottom ).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin

    doi: 10.1073/pnas.182372299

    Figure Lengend Snippet: Expression of TRX in transformed cells cultured with SAHA. T24 bladder carcinoma cells were cultured with vehicle alone (0) or 2.5 or 5 μM SAHA for 6, 15, or 24 h. RNA was extracted and analyzed by Northern blotting for levels of TRX, using a 500-bp 32 P-labeled cDNA probe ( Top ). The blots were subsequently rehybridized with the 1.1-kb 32 P-labeled TBP-2 cDNA probe to confirm induction of TBP-2 ( Middle ) and a γ- 32 P-labeled 18S oligonucleotide probe to indicate RNA loading ( Bottom ).

    Article Snippet: Rapid amplification of cDNA ends was performed to determine the transcriptional start site of the TBP-2 gene, using TBP-2-specific primer 1 (5′-GTTGGTTTTAAGAGTTAGAAATGACGG-3′) and nested primer 2 (5′-TAAGGTATTCTTAAGCAGTTTGAGC-3′) with Marathon-ready human brain cDNA (CLONTECH), according to the manufacturer's instructions.

    Techniques: Expressing, Transformation Assay, Cell Culture, Northern Blot, Labeling

    Induction of TBP-2 mRNA levels in transformed cells by SAHA. LNCaP prostate carcinoma and T24 bladder carcinoma cells were cultured with vehicle alone (0) or SAHA (2.5 or 7.5 μM) for the indicated times. Total RNA was extracted from the cells, and the levels of TBP-2 were determined by Northern blotting using a 1.1-kb 32 P-labeled TBP-2 cDNA probe ( Upper for each cell line). Blots were rehybridized with a γ- 32 P-labeled 18S oligonucleotide probe to indicate RNA loading and are shown ( Lower for each cell line). Similar results were obtained for a total of six transformed cell lines.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin

    doi: 10.1073/pnas.182372299

    Figure Lengend Snippet: Induction of TBP-2 mRNA levels in transformed cells by SAHA. LNCaP prostate carcinoma and T24 bladder carcinoma cells were cultured with vehicle alone (0) or SAHA (2.5 or 7.5 μM) for the indicated times. Total RNA was extracted from the cells, and the levels of TBP-2 were determined by Northern blotting using a 1.1-kb 32 P-labeled TBP-2 cDNA probe ( Upper for each cell line). Blots were rehybridized with a γ- 32 P-labeled 18S oligonucleotide probe to indicate RNA loading and are shown ( Lower for each cell line). Similar results were obtained for a total of six transformed cell lines.

    Article Snippet: Rapid amplification of cDNA ends was performed to determine the transcriptional start site of the TBP-2 gene, using TBP-2-specific primer 1 (5′-GTTGGTTTTAAGAGTTAGAAATGACGG-3′) and nested primer 2 (5′-TAAGGTATTCTTAAGCAGTTTGAGC-3′) with Marathon-ready human brain cDNA (CLONTECH), according to the manufacturer's instructions.

    Techniques: Transformation Assay, Cell Culture, Northern Blot, Labeling

    Annotated variants of the GRN 5′ UTR. A , sequence of the 219 nucleotides of the GRN 5′ UTR derived from human brain cDNA. This annotated variant ( NM002087 ) and start sites of predicted shorter variants ( XM005257255 and XM005257254 ) are indicated, as well as a predicted splice variant ( XM005257253 ) containing a short sequence stretch of exon 1, followed by an insertion of 271 nucleotides of intron 1 ( green box ) at the exon 1/2 boundary. 5′ ends identified by rapid amplification of cDNA ends are marked by arrows. Green arrows indicate mRNAs containing the intron 1-derived sequence stretch. ORFs are highlighted in red , and AUG start codons are circled. B , schematic of the 219-nucleotide-long GRN 5′ UTR. Two AUG initiation codons (positions 90 and 120), the corresponding uORF, and the stop codon (nucleotides 159–161) are indicated. C , schematic of the alternative spliced GRN 5′ UTR ( XM005257253 ). This splice variant contains a short stretch of exon 1 (nucleotides 190–212), followed by 271 nucleotides derived from intron 1 containing two AUGs ( circled ) and one short uORF ( 156–176 ).

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: Annotated variants of the GRN 5′ UTR. A , sequence of the 219 nucleotides of the GRN 5′ UTR derived from human brain cDNA. This annotated variant ( NM002087 ) and start sites of predicted shorter variants ( XM005257255 and XM005257254 ) are indicated, as well as a predicted splice variant ( XM005257253 ) containing a short sequence stretch of exon 1, followed by an insertion of 271 nucleotides of intron 1 ( green box ) at the exon 1/2 boundary. 5′ ends identified by rapid amplification of cDNA ends are marked by arrows. Green arrows indicate mRNAs containing the intron 1-derived sequence stretch. ORFs are highlighted in red , and AUG start codons are circled. B , schematic of the 219-nucleotide-long GRN 5′ UTR. Two AUG initiation codons (positions 90 and 120), the corresponding uORF, and the stop codon (nucleotides 159–161) are indicated. C , schematic of the alternative spliced GRN 5′ UTR ( XM005257253 ). This splice variant contains a short stretch of exon 1 (nucleotides 190–212), followed by 271 nucleotides derived from intron 1 containing two AUGs ( circled ) and one short uORF ( 156–176 ).

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Sequencing, Derivative Assay, Variant Assay, Rapid Amplification of cDNA Ends

    Inhibition of GRN expression occurs at the translational and mRNA levels. A , mRNAs encoding the indicated GRN variants were transcribed in vitro. B , equal amounts of in vitro transcribed mRNAs (∼1 μg, adjusted to the number of nucleotides per individual mRNA) were used for in vitro translation. Note that the 3′ UTR does not affect GRN translation, whereas the 5′ UTR strongly represses GRN translation. Bottom panel , quantitation of the expression levels of GRN upon in vitro translation of equal amounts of mRNA normalized to expression of the 5′3′ UTR cDNA construct. C , quantitation of GRN mRNA in stable cell lines expressing the indicated cDNA constructs. Levels are normalized to expression of the 5′3′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: Inhibition of GRN expression occurs at the translational and mRNA levels. A , mRNAs encoding the indicated GRN variants were transcribed in vitro. B , equal amounts of in vitro transcribed mRNAs (∼1 μg, adjusted to the number of nucleotides per individual mRNA) were used for in vitro translation. Note that the 3′ UTR does not affect GRN translation, whereas the 5′ UTR strongly represses GRN translation. Bottom panel , quantitation of the expression levels of GRN upon in vitro translation of equal amounts of mRNA normalized to expression of the 5′3′ UTR cDNA construct. C , quantitation of GRN mRNA in stable cell lines expressing the indicated cDNA constructs. Levels are normalized to expression of the 5′3′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). *, p

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Inhibition, Expressing, In Vitro, Quantitation Assay, Construct, Stable Transfection

    GRN expression is inhibited by its 5 ′ -UTR. A , cDNA constructs used to evaluate the effects of the 5′ and the 3′ UTR on GRN expression. B , isogenic expression of the cDNA constructs shown in A in HEK 293 cells. GRN expression was investigated in cell lysates by Western blotting using an anti-myc or anti-GRN antibody. Actin was used as loading control. Quantitation of the expression levels of GRN in cell lysates and of secreted GRN is shown in the bottom panel ). For all quantifications, data are mean ± S.D. ( n ≥ 3 independent experiments). ****, p

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: GRN expression is inhibited by its 5 ′ -UTR. A , cDNA constructs used to evaluate the effects of the 5′ and the 3′ UTR on GRN expression. B , isogenic expression of the cDNA constructs shown in A in HEK 293 cells. GRN expression was investigated in cell lysates by Western blotting using an anti-myc or anti-GRN antibody. Actin was used as loading control. Quantitation of the expression levels of GRN in cell lysates and of secreted GRN is shown in the bottom panel ). For all quantifications, data are mean ± S.D. ( n ≥ 3 independent experiments). ****, p

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Expressing, Construct, Western Blot, Quantitation Assay

    AUGs in the uORF are responsible for translational repression of GRN but not for reducing mRNA levels. A , schematic of the investigated cDNA constructs. B , isogenic expression of the cDNA constructs shown in A . Actin was used to verify equal loading. Bottom panel , quantitation of the expression levels of the GRN variants normalized to the 5′ UTR. Levels are normalized to expression of the 5′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). ***, p

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: AUGs in the uORF are responsible for translational repression of GRN but not for reducing mRNA levels. A , schematic of the investigated cDNA constructs. B , isogenic expression of the cDNA constructs shown in A . Actin was used to verify equal loading. Bottom panel , quantitation of the expression levels of the GRN variants normalized to the 5′ UTR. Levels are normalized to expression of the 5′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). ***, p

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Construct, Expressing, Quantitation Assay

    Both AUGs in the uORF can initiate translation. A , schematic of the uORF in-frame with EGFP containing a mutated start codon. The two mutated uORF start codons are indicated ( AUA 1 and AUA 2 ). B , transient expression of EGFP cDNA or cDNA constructs containing nucleotides 76–158 of the 5′ UTR including both AUGs ( 76–158 ), either one ( 76–158 AUA1 and 76–158 AUA2 ), or both ( 76–158 AUA1,2 ) mutated AUGs. Actin was used as a loading control. Note that both start codons of the uORF are functional because they are capable to initiate protein translation. The top and bottom bands represent proteins translated from the first and second AUG of the uORF. When both AUGs are mutated, no translation product can be detected.

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: Both AUGs in the uORF can initiate translation. A , schematic of the uORF in-frame with EGFP containing a mutated start codon. The two mutated uORF start codons are indicated ( AUA 1 and AUA 2 ). B , transient expression of EGFP cDNA or cDNA constructs containing nucleotides 76–158 of the 5′ UTR including both AUGs ( 76–158 ), either one ( 76–158 AUA1 and 76–158 AUA2 ), or both ( 76–158 AUA1,2 ) mutated AUGs. Actin was used as a loading control. Note that both start codons of the uORF are functional because they are capable to initiate protein translation. The top and bottom bands represent proteins translated from the first and second AUG of the uORF. When both AUGs are mutated, no translation product can be detected.

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Expressing, Construct, Functional Assay

    A 50-nucleotide stretch within the 5′ UTR of GRN is required for translational inhibition and lowering of mRNA levels. A , schematic of investigated cDNA constructs. B , deletion of nucleotides 75–125 (Δ 75–125 ) is sufficient to abolish repression of GRN expression, whereas mutagenesis of both in-frame AUGs within the 5′ UTR increases GRN expression to a significantly lower extent. Bottom panel , quantitation of the expression levels of the GRN variants shown in the top panel . Expression is normalized to the 5′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: A 50-nucleotide stretch within the 5′ UTR of GRN is required for translational inhibition and lowering of mRNA levels. A , schematic of investigated cDNA constructs. B , deletion of nucleotides 75–125 (Δ 75–125 ) is sufficient to abolish repression of GRN expression, whereas mutagenesis of both in-frame AUGs within the 5′ UTR increases GRN expression to a significantly lower extent. Bottom panel , quantitation of the expression levels of the GRN variants shown in the top panel . Expression is normalized to the 5′ UTR cDNA construct. Data are mean ± S.D. ( n ≥ 3 independent experiments). *, p

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Inhibition, Construct, Expressing, Mutagenesis, Quantitation Assay

    The GRN 5′ UTR inhibits expression of an EGFP reporter. A , schematic showing cDNA constructs used to evaluate the effects of the 5′ UTR, 3′ UTR, and both 5′3′ UTR of GRN on EGFP expression. To allow secretion of EGFP, the coding sequence of EGFP was fused to the signal peptide ( SP ) derived from GRN ( SP EGFP ). B , isogenic expression of SP EGFP variants with and without the GRN 5′ UTR, 3′ UTR, and 5′3′ UTR. Actin was used as a loading control. Bottom panel , expression levels of SP EGFP in cell lysates and corresponding media were quantified. All quantifications were normalized to 5′3′ UTR constructs. Data are mean ± S.D. ( n ≥ 3 independent experiments). **, p

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin Transcripts with Short and Long 5′ Untranslated Regions (UTRs) Are Differentially Expressed via Posttranscriptional and Translational Repression *

    doi: 10.1074/jbc.M114.560128

    Figure Lengend Snippet: The GRN 5′ UTR inhibits expression of an EGFP reporter. A , schematic showing cDNA constructs used to evaluate the effects of the 5′ UTR, 3′ UTR, and both 5′3′ UTR of GRN on EGFP expression. To allow secretion of EGFP, the coding sequence of EGFP was fused to the signal peptide ( SP ) derived from GRN ( SP EGFP ). B , isogenic expression of SP EGFP variants with and without the GRN 5′ UTR, 3′ UTR, and 5′3′ UTR. Actin was used as a loading control. Bottom panel , expression levels of SP EGFP in cell lysates and corresponding media were quantified. All quantifications were normalized to 5′3′ UTR constructs. Data are mean ± S.D. ( n ≥ 3 independent experiments). **, p

    Article Snippet: Furthermore, the 219-nucleotide-long 5′ UTR could be amplified by PCR from an adult human brain cDNA (OriGene Technologies, Rockville, MD) (data not shown).

    Techniques: Expressing, Construct, Sequencing, Derivative Assay

    Expression Studies of SYT14/Syt14 cDNA in Human and Mouse Tissues and Localization of SYT14 in Transfected COS-1 Cells

    Journal: American Journal of Human Genetics

    Article Title: Exome Sequencing Reveals a Homozygous SYT14 Mutation in Adult-Onset, Autosomal-Recessive Spinocerebellar Ataxia with Psychomotor Retardation

    doi: 10.1016/j.ajhg.2011.07.012

    Figure Lengend Snippet: Expression Studies of SYT14/Syt14 cDNA in Human and Mouse Tissues and Localization of SYT14 in Transfected COS-1 Cells

    Article Snippet: The full-length SYT14 PCR product amplified from human brain cDNA (MHS4426-99239810, Open Biosystems, Huntsville, AL) was used as a template and subcloned into pDONR221 (the entry vector of Gateway system, Invitrogen).

    Techniques: Expressing, Transfection