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  • 91
    Thermo Fisher region flanking rs27324996
    Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of <t>rs27324996,</t> Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.
    Region Flanking Rs27324996, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    region flanking rs27324996 - by Bioz Stars, 2020-08
    91/100 stars
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    85
    Aalto Bio Reagents c5 region
    Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of <t>rs27324996,</t> Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.
    C5 Region, supplied by Aalto Bio Reagents, used in various techniques. Bioz Stars score: 85/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c5 region/product/Aalto Bio Reagents
    Average 85 stars, based on 13 article reviews
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    c5 region - by Bioz Stars, 2020-08
    85/100 stars
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    91
    Santa Cruz Biotechnology eiti region
    Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of <t>rs27324996,</t> Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.
    Eiti Region, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/eiti region/product/Santa Cruz Biotechnology
    Average 91 stars, based on 2 article reviews
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    eiti region - by Bioz Stars, 2020-08
    91/100 stars
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    85
    Northern California Research geographical region
    Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of <t>rs27324996,</t> Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.
    Geographical Region, supplied by Northern California Research, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/geographical region/product/Northern California Research
    Average 85 stars, based on 2 article reviews
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    geographical region - by Bioz Stars, 2020-08
    85/100 stars
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    88
    Syntaxin juxtamembrane region
    In vivo effects of single, double, and triple point mutations in the Sso1p <t>juxtamembrane</t> region. A. Growth on 5-FOA with single point mutation. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY384 (Sso1p-A259G-HA), JMY380 (Sso1p-R260G-HA), JMY381 (Sso1p-K261G-HA), JMY382 (Sso1p-K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY394 (Sso1p-A259G-HA), JMY395 (Sso1p-R260G-HA), JMY396 (Sso1p-K261G-HA), JMY397 (Sso1p-K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. B. Growth on 5-FOA with double and triple point mutations. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY376 (Sso1p-R260G, K261G-HA), JMY377 (Sso1p-R260G, K263GHA), JMY378 (Sso1p-K261G, K263G-HA), JMY379 (Sso1p-R260G, K261G, K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY398 (Sso1p-R260G, K261G-HA), JMY399 (Sso1p-R260G, K263G-HA), JMY400 (Sso1p-K261G, K263G-HA), JMY401 (Sso1p-R260G, K261G, K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h.
    Juxtamembrane Region, supplied by Syntaxin, used in various techniques. Bioz Stars score: 88/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    juxtamembrane region - by Bioz Stars, 2020-08
    88/100 stars
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    89
    Roche ns5b region
    In vivo effects of single, double, and triple point mutations in the Sso1p <t>juxtamembrane</t> region. A. Growth on 5-FOA with single point mutation. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY384 (Sso1p-A259G-HA), JMY380 (Sso1p-R260G-HA), JMY381 (Sso1p-K261G-HA), JMY382 (Sso1p-K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY394 (Sso1p-A259G-HA), JMY395 (Sso1p-R260G-HA), JMY396 (Sso1p-K261G-HA), JMY397 (Sso1p-K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. B. Growth on 5-FOA with double and triple point mutations. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY376 (Sso1p-R260G, K261G-HA), JMY377 (Sso1p-R260G, K263GHA), JMY378 (Sso1p-K261G, K263G-HA), JMY379 (Sso1p-R260G, K261G, K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY398 (Sso1p-R260G, K261G-HA), JMY399 (Sso1p-R260G, K263G-HA), JMY400 (Sso1p-K261G, K263G-HA), JMY401 (Sso1p-R260G, K261G, K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h.
    Ns5b Region, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ns5b region/product/Roche
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    ns5b region - by Bioz Stars, 2020-08
    89/100 stars
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    91
    Gallus BioPharmaceuticals pg1 region
    Structural analysis of p190RhoGAP pseudoGTPase domain <t>pG1.</t> a Crystal structures of pG1 for p190RhoGAP-A ( left , cyan ) and p190RhoGAP-B ( right , blue ). Secondary structure and N and C termini are labeled. Unmodeled loops are indicated by a dashed line . b Superposition of p190RhoGAP-A and -B pG1 domains. c Superposition of p190RhoGAP-A pG1 domain with GTP-analog bound H-Ras in gray ; PDB ID: 5P21 56
    Pg1 Region, supplied by Gallus BioPharmaceuticals, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pg1 region/product/Gallus BioPharmaceuticals
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    pg1 region - by Bioz Stars, 2020-08
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    90
    Abnova polylinker region
    GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the <t>polylinker</t> region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry
    Polylinker Region, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Stratagene protease region
    GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the <t>polylinker</t> region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry
    Protease Region, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    protease region - by Bioz Stars, 2020-08
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    92
    Novo Nordisk region skåne
    GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the <t>polylinker</t> region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry
    Region Skåne, supplied by Novo Nordisk, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    region skåne - by Bioz Stars, 2020-08
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    88
    Pepscan stalk region
    GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the <t>polylinker</t> region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry
    Stalk Region, supplied by Pepscan, 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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    89
    Illumina Inc cnv region
    SNP haplotypes surrounding HP persist through the <t>CNV</t> region, yet segregate with both structural forms of HP This plot displays the SNP haplotypes (10 kb on each side of the HP CNV) segregating with HP1 and HP2 based on an analysis of 264 samples (528 haplotypes). The upstream <t>SNPs</t> are proximal to the centromere, while the downstream SNPs are distal to the centromere. Each thin horizontal line represents an individual SNP haplotype; similar or identical haplotypes are organized into clusters outlined by colored boxes. Note that the size of small clusters has been increased for visibility purposes and the number of haplotypes contained in each cluster is indicated at the left of the plot. White represents the minor allele and grey indicates the major allele across all populations in the analysis (CEU, IBS, TSI, YRI). Haplotypes ascertained from West African (HapMap YRI) individuals are indicated with lavender bars to the left of the plot, while haplotypes ascertained in European populations (CEU, IBS, TSI) are indicated with dark purple bars to the left of the plot. Haplotypes were clustered with the k-means method using upstream SNP haplotypes. Similar SNP haplotypes carrying different structures are indicated with colored outlines (dark pink, light blue, green, gold) and are designated haplotypes A–D. This figure was based on analysis of 1,000 Genomes Project samples and data (Methods).
    Cnv Region, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 89/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cnv region - by Bioz Stars, 2020-08
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    90
    Syntaxin cytoplasmic region
    SNP haplotypes surrounding HP persist through the <t>CNV</t> region, yet segregate with both structural forms of HP This plot displays the SNP haplotypes (10 kb on each side of the HP CNV) segregating with HP1 and HP2 based on an analysis of 264 samples (528 haplotypes). The upstream <t>SNPs</t> are proximal to the centromere, while the downstream SNPs are distal to the centromere. Each thin horizontal line represents an individual SNP haplotype; similar or identical haplotypes are organized into clusters outlined by colored boxes. Note that the size of small clusters has been increased for visibility purposes and the number of haplotypes contained in each cluster is indicated at the left of the plot. White represents the minor allele and grey indicates the major allele across all populations in the analysis (CEU, IBS, TSI, YRI). Haplotypes ascertained from West African (HapMap YRI) individuals are indicated with lavender bars to the left of the plot, while haplotypes ascertained in European populations (CEU, IBS, TSI) are indicated with dark purple bars to the left of the plot. Haplotypes were clustered with the k-means method using upstream SNP haplotypes. Similar SNP haplotypes carrying different structures are indicated with colored outlines (dark pink, light blue, green, gold) and are designated haplotypes A–D. This figure was based on analysis of 1,000 Genomes Project samples and data (Methods).
    Cytoplasmic Region, supplied by Syntaxin, used in various techniques. Bioz Stars score: 90/100, based on 141 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cytoplasmic region/product/Syntaxin
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    cytoplasmic region - by Bioz Stars, 2020-08
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    86
    Epitomics npty region
    SNP haplotypes surrounding HP persist through the <t>CNV</t> region, yet segregate with both structural forms of HP This plot displays the SNP haplotypes (10 kb on each side of the HP CNV) segregating with HP1 and HP2 based on an analysis of 264 samples (528 haplotypes). The upstream <t>SNPs</t> are proximal to the centromere, while the downstream SNPs are distal to the centromere. Each thin horizontal line represents an individual SNP haplotype; similar or identical haplotypes are organized into clusters outlined by colored boxes. Note that the size of small clusters has been increased for visibility purposes and the number of haplotypes contained in each cluster is indicated at the left of the plot. White represents the minor allele and grey indicates the major allele across all populations in the analysis (CEU, IBS, TSI, YRI). Haplotypes ascertained from West African (HapMap YRI) individuals are indicated with lavender bars to the left of the plot, while haplotypes ascertained in European populations (CEU, IBS, TSI) are indicated with dark purple bars to the left of the plot. Haplotypes were clustered with the k-means method using upstream SNP haplotypes. Similar SNP haplotypes carrying different structures are indicated with colored outlines (dark pink, light blue, green, gold) and are designated haplotypes A–D. This figure was based on analysis of 1,000 Genomes Project samples and data (Methods).
    Npty Region, supplied by Epitomics, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/npty region/product/Epitomics
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    npty region - by Bioz Stars, 2020-08
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    93
    Illumina Inc v4 region
    Alignment of isolated strain 16S rRNA <t>V4</t> region and representative sequence of Lactobacillus in metagenomics.
    V4 Region, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 93/100, based on 3008 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 3008 article reviews
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    v4 region - by Bioz Stars, 2020-08
    93/100 stars
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    92
    BioLegend aβ region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Aβ Region, supplied by BioLegend, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    aβ region - by Bioz Stars, 2020-08
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    93
    Vital River Laboratories dorsal region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
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    Santa Cruz Biotechnology meridional region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Meridional Region, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Genechem utr region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
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    5 PRIME barcoding region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
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    89
    Roche interscapular region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Interscapular Region, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SwitchGear Genomics nongenic region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Nongenic Region, supplied by SwitchGear Genomics, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    4Gene upstream region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Upstream Region, supplied by 4Gene, used in various techniques. Bioz Stars score: 90/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene utr region
    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of <t>Aβ</t> and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice
    Utr Region, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eurofins v4 region
    <t>V4</t> region and 5′ and 3′ PCR primers. The target nucleotides for the PCR primers are depicted in grey (5′region) and black (3′region). The 5′ and 3′ primers include adaptor Illumina sequences for NexGen sequences
    V4 Region, supplied by Eurofins, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of rs27324996, Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.

    Journal: Cell stem cell

    Article Title: Large diverse population cohorts of hiPSCs and derived hepatocyte-like cells reveal functional genetic variation at blood lipid-associated loci

    doi: 10.1016/j.stem.2017.03.017

    Figure Lengend Snippet: Evidence for rs2277862- CPNE1 as a Functional SNP-Gene Set (A) Schematics of human chromosome 20q11 locus showing the relative positions of rs2277862, CPNE1 , and ERGIC3 and mouse chromosome 2qH1 locus showing the relative positions of rs27324996, Cpne1 , and Ergic3 . (B) Top panels: heterozygous knock-in of rs2277862 minor allele with a single-strand DNA oligonucleotide. Representative indels in non-knock-in clones are also shown. Bottom panels: homozygous 38-bp deletions (“knockout” or Δ38/Δ38) encompassing rs2277862 using a dual gRNA approach. A representative agarose gel of PCR amplicons is shown. The protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. (C) Left panels: gene expression in undifferentiated HUES 8 cells (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) and differentiated HUES 8 HLCs (n = 2 wild-type clones and 1 knock-in clone; 6 wells per clone) normalized to mean expression level in wild-type clones. Right panels: gene expression in undifferentiated HUES 8 cells (n = 10 wild-type and 10 knockout clones, 3 wells per clone) and undifferentiated H7 cells (n = 8 wild-type and 6 knockout clones, 3 wells per clone) normalized to mean expression level in wild-type clones. (D) CRISPR interference at the rs2277862 site. The three gRNA protospacers are underlined, the PAMs are bolded, and the SNP position is indicated in red. The graphs show gene expression, normalized to mean expression level in control cells, in HEK 293T cells transfected with catalytically dead Cas9 (dCas9) with the gRNAs (either singly or in combination, 3 wells per condition). Control cells were transfected with dCas9 without an accompanying gRNA. (E) The noncoding rs2277862 site is well conserved in mouse, including allelic variants of the SNP itself, with the murine equivalent being rs27324996. The SNP position is indicated in red, non-conserved nucleotides are indicated in blue, the gRNA protospacer used to generate the knock-in mouse is underlined, and the PAM is bolded. The electropherogram is from a mouse in which the minor allele of rs2277862/rs27324996 (T) has been knocked into one chromosome, along with four non-conserved nucleotides to “humanize” the site. (F) Gene expression in liver from littermates of the C57BL/6J background (n = 18 wild-type mice and 10 homozygous knock-in mice), normalized to mean expression level in wild-type mice. Data are displayed as means and s.e.m. P -values were calculated with two-tailed Welch’s t -tests.

    Article Snippet: The region flanking rs27324996 was PCR amplified (F: 5′-TGGGAATGGCTTCTTAGGGC-3′ and R: 5′-CATCCCCAAGCAACTCAACC-3′) using AccuPrime Taq DNA Polymerase (Thermo Fisher Scientific) with the following cycling conditions: 94°C for 2 min, [94°C for 30 sec, 55°C for 30 sec, 68°C for 30 sec] × 40 cycles, 68°C for 5 min. PCR products were purified using the DNA Clean & Concentrator kit (Zymo Research) and analyzed for the presence of indels using the Surveyor Mutation Detection Kit (Integrated DNA Technologies) according to the manufacturer’s instructions.

    Techniques: Functional Assay, Knock-In, Clone Assay, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Expressing, Knock-Out, CRISPR, Transfection, Mouse Assay, Two Tailed Test

    In vivo effects of single, double, and triple point mutations in the Sso1p juxtamembrane region. A. Growth on 5-FOA with single point mutation. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY384 (Sso1p-A259G-HA), JMY380 (Sso1p-R260G-HA), JMY381 (Sso1p-K261G-HA), JMY382 (Sso1p-K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY394 (Sso1p-A259G-HA), JMY395 (Sso1p-R260G-HA), JMY396 (Sso1p-K261G-HA), JMY397 (Sso1p-K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. B. Growth on 5-FOA with double and triple point mutations. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY376 (Sso1p-R260G, K261G-HA), JMY377 (Sso1p-R260G, K263GHA), JMY378 (Sso1p-K261G, K263G-HA), JMY379 (Sso1p-R260G, K261G, K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY398 (Sso1p-R260G, K261G-HA), JMY399 (Sso1p-R260G, K263G-HA), JMY400 (Sso1p-K261G, K263G-HA), JMY401 (Sso1p-R260G, K261G, K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h.

    Journal: Eukaryotic Cell

    Article Title: The Polybasic Juxtamembrane Region of Sso1p Is Required for SNARE Function In Vivo †

    doi: 10.1128/EC.4.12.2017-2028.2005

    Figure Lengend Snippet: In vivo effects of single, double, and triple point mutations in the Sso1p juxtamembrane region. A. Growth on 5-FOA with single point mutation. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY384 (Sso1p-A259G-HA), JMY380 (Sso1p-R260G-HA), JMY381 (Sso1p-K261G-HA), JMY382 (Sso1p-K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY394 (Sso1p-A259G-HA), JMY395 (Sso1p-R260G-HA), JMY396 (Sso1p-K261G-HA), JMY397 (Sso1p-K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. B. Growth on 5-FOA with double and triple point mutations. Left: threefold serial dilutions of JMY303 (Sso1p-HA), JMY376 (Sso1p-R260G, K261G-HA), JMY377 (Sso1p-R260G, K263GHA), JMY378 (Sso1p-K261G, K263G-HA), JMY379 (Sso1p-R260G, K261G, K263G-HA), or JMY305 (Sso1pΔNRD) were spotted onto synthetic complete media with 2% galactose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h. Right: threefold serial dilutions of JMY384 (Sso1p-HA), JMY398 (Sso1p-R260G, K261G-HA), JMY399 (Sso1p-R260G, K263G-HA), JMY400 (Sso1p-K261G, K263G-HA), JMY401 (Sso1p-R260G, K261G, K263G-HA), or JMY402 (vector) were spotted onto synthetic complete media with 2% glucose containing 1 mg/ml 5-fluoroorotic acid and grown at 30°C for 72 h.

    Article Snippet: These mutant proteins address two related questions with respect to the distance between the core complex domain and the transmembrane domain: is the distance between the core complex domain and the transmembrane domain critical for function, and what degree of flexibility is permitted in the juxtamembrane region to maintain function?

    Techniques: In Vivo, Mutagenesis, Plasmid Preparation

    In vitro fusion reactions with recombinant Sso1p juxtamembrane insertions. A. Kinetic fusion assay comparing different acceptor t-SNARE liposomes containing t-SNARE complexes composed of GST-Sec9c and the indicated Sso1 protein. Each t-SNARE liposome population (45 μl) was mixed with fluorescent donor v-SNARE liposomes containing Snc1p (5 μl) and NBD fluorescence monitored in a fluorescent plate reader for 2 h. B. Coomassie blue-stained gel of the liposomes used in panel A, indicating that very similar amounts of various Sso1p mutants were reconstituted.

    Journal: Eukaryotic Cell

    Article Title: The Polybasic Juxtamembrane Region of Sso1p Is Required for SNARE Function In Vivo †

    doi: 10.1128/EC.4.12.2017-2028.2005

    Figure Lengend Snippet: In vitro fusion reactions with recombinant Sso1p juxtamembrane insertions. A. Kinetic fusion assay comparing different acceptor t-SNARE liposomes containing t-SNARE complexes composed of GST-Sec9c and the indicated Sso1 protein. Each t-SNARE liposome population (45 μl) was mixed with fluorescent donor v-SNARE liposomes containing Snc1p (5 μl) and NBD fluorescence monitored in a fluorescent plate reader for 2 h. B. Coomassie blue-stained gel of the liposomes used in panel A, indicating that very similar amounts of various Sso1p mutants were reconstituted.

    Article Snippet: These mutant proteins address two related questions with respect to the distance between the core complex domain and the transmembrane domain: is the distance between the core complex domain and the transmembrane domain critical for function, and what degree of flexibility is permitted in the juxtamembrane region to maintain function?

    Techniques: In Vitro, Recombinant, Single Vesicle Fusion Assay, Fluorescence, Staining

    Structural analysis of p190RhoGAP pseudoGTPase domain pG1. a Crystal structures of pG1 for p190RhoGAP-A ( left , cyan ) and p190RhoGAP-B ( right , blue ). Secondary structure and N and C termini are labeled. Unmodeled loops are indicated by a dashed line . b Superposition of p190RhoGAP-A and -B pG1 domains. c Superposition of p190RhoGAP-A pG1 domain with GTP-analog bound H-Ras in gray ; PDB ID: 5P21 56

    Journal: Nature Communications

    Article Title: p190RhoGAP proteins contain pseudoGTPase domains

    doi: 10.1038/s41467-017-00483-x

    Figure Lengend Snippet: Structural analysis of p190RhoGAP pseudoGTPase domain pG1. a Crystal structures of pG1 for p190RhoGAP-A ( left , cyan ) and p190RhoGAP-B ( right , blue ). Secondary structure and N and C termini are labeled. Unmodeled loops are indicated by a dashed line . b Superposition of p190RhoGAP-A and -B pG1 domains. c Superposition of p190RhoGAP-A pG1 domain with GTP-analog bound H-Ras in gray ; PDB ID: 5P21 56

    Article Snippet: Codon-optimized synthetic cDNAs (Supplementary Table ) encoding the pG1 region from Gallus gallus (chicken) p190RhoGAP-A (UniProt ID: A0A1D5P6Q7, residues 592–764, 90% identical to rat p190RhoGAP-A), Xenopus laevis (frog) p190RhoGAP-A (UniProt ID: Q6NU25, residues 587–762, 89% identical to rat p190RhoGAP-A), Danio rerio (zebrafish) p190RhoGAP-A (UniProt ID: F1R0X6, residues 592–772, 69% identical to rat p190RhoGAP-A) and D. melanogaster p190RhoGAP, which contains a single gene for p190 (UniProt ID: Q9VX32, residues 593–753, 22% identical to rat p190RhoGAP-A), were purchased from GenScript.

    Techniques: Labeling

    Assessment of nucleotide binding by p190RhoGAP pG1. a – c Fluorescence of MANT-GTPγS monitored over time upon addition of proteins as indicated. a pG1 from 190RhoGAP-A or -B, or Rac1 as a control, in the presence of 20 mM EDTA plus 10 mM MgCl 2 . b p190RhoGAP-A pG1 of Xenopus laevis (frog), Gallus gallus (chicken) and Danio rerio (zebrafish) or the single p190RhoGAP of Drosophila melanogaster . c p190RhoGAP-A pG1 in the presence of different divalent cations. Rac1 in Mg 2+ is included as a positive control. Uncorrected for photobleaching. d – i Thermal shift assay. d – f Thermal denaturation curves (from a representative experiment) of pG1 domains or Rac1 in the presence of divalent cation, nucleotide or both as listed in f . Buffer alone curves ( black ) are labeled with arrows . g – i Histograms of melting temperature changes (Δ T m ) compared to buffer alone, determined by fitting the thermal denaturation curves ( d – f ) to a sigmoidal model. Error bars indicate s.e.m. ( n =3). Positive shifts indicate a stabilization in the presence of ligand

    Journal: Nature Communications

    Article Title: p190RhoGAP proteins contain pseudoGTPase domains

    doi: 10.1038/s41467-017-00483-x

    Figure Lengend Snippet: Assessment of nucleotide binding by p190RhoGAP pG1. a – c Fluorescence of MANT-GTPγS monitored over time upon addition of proteins as indicated. a pG1 from 190RhoGAP-A or -B, or Rac1 as a control, in the presence of 20 mM EDTA plus 10 mM MgCl 2 . b p190RhoGAP-A pG1 of Xenopus laevis (frog), Gallus gallus (chicken) and Danio rerio (zebrafish) or the single p190RhoGAP of Drosophila melanogaster . c p190RhoGAP-A pG1 in the presence of different divalent cations. Rac1 in Mg 2+ is included as a positive control. Uncorrected for photobleaching. d – i Thermal shift assay. d – f Thermal denaturation curves (from a representative experiment) of pG1 domains or Rac1 in the presence of divalent cation, nucleotide or both as listed in f . Buffer alone curves ( black ) are labeled with arrows . g – i Histograms of melting temperature changes (Δ T m ) compared to buffer alone, determined by fitting the thermal denaturation curves ( d – f ) to a sigmoidal model. Error bars indicate s.e.m. ( n =3). Positive shifts indicate a stabilization in the presence of ligand

    Article Snippet: Codon-optimized synthetic cDNAs (Supplementary Table ) encoding the pG1 region from Gallus gallus (chicken) p190RhoGAP-A (UniProt ID: A0A1D5P6Q7, residues 592–764, 90% identical to rat p190RhoGAP-A), Xenopus laevis (frog) p190RhoGAP-A (UniProt ID: Q6NU25, residues 587–762, 89% identical to rat p190RhoGAP-A), Danio rerio (zebrafish) p190RhoGAP-A (UniProt ID: F1R0X6, residues 592–772, 69% identical to rat p190RhoGAP-A) and D. melanogaster p190RhoGAP, which contains a single gene for p190 (UniProt ID: Q9VX32, residues 593–753, 22% identical to rat p190RhoGAP-A), were purchased from GenScript.

    Techniques: Binding Assay, Fluorescence, Positive Control, Thermal Shift Assay, Labeling

    Analysis of the degraded nucleotide binding pocket of p190RhoGAP pG1. a Alignment ( top ) and structural comparison ( bottom ) of conserved G motifs for H-Ras (PDB ID: 5P21 56 ) and p190RhoGAP-A pG1 (p190A). Conserved threonine T35 for H-Ras is shown. Dashed box indicates region shown in b . b Close-up of the disrupted nucleotide-binding site of p190RhoGAP-A pG1 domain ( left ) and additional space filling model indicating the location of GTP analog when H-Ras is superposed onto the structure ( right )

    Journal: Nature Communications

    Article Title: p190RhoGAP proteins contain pseudoGTPase domains

    doi: 10.1038/s41467-017-00483-x

    Figure Lengend Snippet: Analysis of the degraded nucleotide binding pocket of p190RhoGAP pG1. a Alignment ( top ) and structural comparison ( bottom ) of conserved G motifs for H-Ras (PDB ID: 5P21 56 ) and p190RhoGAP-A pG1 (p190A). Conserved threonine T35 for H-Ras is shown. Dashed box indicates region shown in b . b Close-up of the disrupted nucleotide-binding site of p190RhoGAP-A pG1 domain ( left ) and additional space filling model indicating the location of GTP analog when H-Ras is superposed onto the structure ( right )

    Article Snippet: Codon-optimized synthetic cDNAs (Supplementary Table ) encoding the pG1 region from Gallus gallus (chicken) p190RhoGAP-A (UniProt ID: A0A1D5P6Q7, residues 592–764, 90% identical to rat p190RhoGAP-A), Xenopus laevis (frog) p190RhoGAP-A (UniProt ID: Q6NU25, residues 587–762, 89% identical to rat p190RhoGAP-A), Danio rerio (zebrafish) p190RhoGAP-A (UniProt ID: F1R0X6, residues 592–772, 69% identical to rat p190RhoGAP-A) and D. melanogaster p190RhoGAP, which contains a single gene for p190 (UniProt ID: Q9VX32, residues 593–753, 22% identical to rat p190RhoGAP-A), were purchased from GenScript.

    Techniques: Binding Assay

    Rho activation assays. a Active endogenous RhoA levels assessed by pulldown with a GST-rhotekin Rho binding domain (GST-RBD) from HEK293T cells transfected with a gradient of Flag-p190RhoGAP-A, pG1-pG2 deletion mutant (ΔpG1–pG2), or empty Flag vector (−). A representative set of immunoblots is shown. b Scatter plot of active RhoA as a function of Flag-p190RhoGAP expression. Active RhoA is normalized to maximum levels in each experiment (empty vector control) and Flag-p190RhoGAP expression as a percentage of maximum expression within each experiment. Shown is the compilation of data from 8 independent experiments. c Scatter/bar plot of data from ( b ), with data points binned according to Flag-p190RhoGAP expression as indicated (1–10%, 10–25%, 25–50% and 50–100%). The mean and s.e.m. ( error bars ) of each group is shown. Unpaired t -tests (two-tailed) were performed in GraphPad Prism, with statistical significance indicated above pairs and p- values as follows: 1–10% bin p =0.0053; 10–25% bin p =0.0019; 25–50% bin p =0.0006; and 50–100% bin p =0.0133

    Journal: Nature Communications

    Article Title: p190RhoGAP proteins contain pseudoGTPase domains

    doi: 10.1038/s41467-017-00483-x

    Figure Lengend Snippet: Rho activation assays. a Active endogenous RhoA levels assessed by pulldown with a GST-rhotekin Rho binding domain (GST-RBD) from HEK293T cells transfected with a gradient of Flag-p190RhoGAP-A, pG1-pG2 deletion mutant (ΔpG1–pG2), or empty Flag vector (−). A representative set of immunoblots is shown. b Scatter plot of active RhoA as a function of Flag-p190RhoGAP expression. Active RhoA is normalized to maximum levels in each experiment (empty vector control) and Flag-p190RhoGAP expression as a percentage of maximum expression within each experiment. Shown is the compilation of data from 8 independent experiments. c Scatter/bar plot of data from ( b ), with data points binned according to Flag-p190RhoGAP expression as indicated (1–10%, 10–25%, 25–50% and 50–100%). The mean and s.e.m. ( error bars ) of each group is shown. Unpaired t -tests (two-tailed) were performed in GraphPad Prism, with statistical significance indicated above pairs and p- values as follows: 1–10% bin p =0.0053; 10–25% bin p =0.0019; 25–50% bin p =0.0006; and 50–100% bin p =0.0133

    Article Snippet: Codon-optimized synthetic cDNAs (Supplementary Table ) encoding the pG1 region from Gallus gallus (chicken) p190RhoGAP-A (UniProt ID: A0A1D5P6Q7, residues 592–764, 90% identical to rat p190RhoGAP-A), Xenopus laevis (frog) p190RhoGAP-A (UniProt ID: Q6NU25, residues 587–762, 89% identical to rat p190RhoGAP-A), Danio rerio (zebrafish) p190RhoGAP-A (UniProt ID: F1R0X6, residues 592–772, 69% identical to rat p190RhoGAP-A) and D. melanogaster p190RhoGAP, which contains a single gene for p190 (UniProt ID: Q9VX32, residues 593–753, 22% identical to rat p190RhoGAP-A), were purchased from GenScript.

    Techniques: Activation Assay, Binding Assay, Transfection, Mutagenesis, Plasmid Preparation, Western Blot, Expressing, Two Tailed Test

    GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the polylinker region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry

    Journal: Cell Death & Disease

    Article Title: NK cell intrinsic regulation of MIP-1α by granzyme M

    doi: 10.1038/cddis.2014.74

    Figure Lengend Snippet: GrzM does not process MIP-1 α . ( a ) Human recombinant full-length GST-tagged MIP-1 α (sequence start mature MIP-1 α : bold letters) was generated. The protein contains a GrzM consensus cleavage site in the polylinker region between GST and the start of full-length MIP-1 α , as well as a potential cleavage site for GrzM in the full-length MIP-1 α sequence. ( b ) This product was then incubated with low concentrations of human recombinant GrzM or a Serine/Alanine (S/A) mutant. The resulting products were used for immunoblotting ( c ) N-terminal sequencing and ( d ) mass spectrometry

    Article Snippet: Western blot analysis Recombinant full-length human GST-tagged MIP-1α , into which a processing consensus site for human GrzM (Valine Leucine Phenylalanine) was encoded in the polylinker region (Abnova, P01), was incubated with human recombinant GrzM or a S/A mutant (inactive) GrzM for the indicated times at 37 °C, or left on ice as a control.

    Techniques: Recombinant, Sequencing, Generated, Incubation, Mutagenesis, Mass Spectrometry

    SNP haplotypes surrounding HP persist through the CNV region, yet segregate with both structural forms of HP This plot displays the SNP haplotypes (10 kb on each side of the HP CNV) segregating with HP1 and HP2 based on an analysis of 264 samples (528 haplotypes). The upstream SNPs are proximal to the centromere, while the downstream SNPs are distal to the centromere. Each thin horizontal line represents an individual SNP haplotype; similar or identical haplotypes are organized into clusters outlined by colored boxes. Note that the size of small clusters has been increased for visibility purposes and the number of haplotypes contained in each cluster is indicated at the left of the plot. White represents the minor allele and grey indicates the major allele across all populations in the analysis (CEU, IBS, TSI, YRI). Haplotypes ascertained from West African (HapMap YRI) individuals are indicated with lavender bars to the left of the plot, while haplotypes ascertained in European populations (CEU, IBS, TSI) are indicated with dark purple bars to the left of the plot. Haplotypes were clustered with the k-means method using upstream SNP haplotypes. Similar SNP haplotypes carrying different structures are indicated with colored outlines (dark pink, light blue, green, gold) and are designated haplotypes A–D. This figure was based on analysis of 1,000 Genomes Project samples and data (Methods).

    Journal: Nature genetics

    Article Title: Recurring exon deletions in the haptoglobin (HP) gene associate with lower blood cholesterol levels

    doi: 10.1038/ng.3510

    Figure Lengend Snippet: SNP haplotypes surrounding HP persist through the CNV region, yet segregate with both structural forms of HP This plot displays the SNP haplotypes (10 kb on each side of the HP CNV) segregating with HP1 and HP2 based on an analysis of 264 samples (528 haplotypes). The upstream SNPs are proximal to the centromere, while the downstream SNPs are distal to the centromere. Each thin horizontal line represents an individual SNP haplotype; similar or identical haplotypes are organized into clusters outlined by colored boxes. Note that the size of small clusters has been increased for visibility purposes and the number of haplotypes contained in each cluster is indicated at the left of the plot. White represents the minor allele and grey indicates the major allele across all populations in the analysis (CEU, IBS, TSI, YRI). Haplotypes ascertained from West African (HapMap YRI) individuals are indicated with lavender bars to the left of the plot, while haplotypes ascertained in European populations (CEU, IBS, TSI) are indicated with dark purple bars to the left of the plot. Haplotypes were clustered with the k-means method using upstream SNP haplotypes. Similar SNP haplotypes carrying different structures are indicated with colored outlines (dark pink, light blue, green, gold) and are designated haplotypes A–D. This figure was based on analysis of 1,000 Genomes Project samples and data (Methods).

    Article Snippet: Imputation of HP structural variation into cohorts for cholesterol association study A reference panel composed of encoded HP structural alleles and SNPs surrounding the CNV region from the Illumina OMNI, Illumina 1M, and Affymetrix 6.0 arrays was developed and used to impute HP structural variation into cohorts with cholesterol information using Beagle (v2.3.1) imputation software.

    Techniques:

    SNP haplotypes and sequence differences between HP subtypes inform structural history (a) This alignment shows base pair differences between HP structural forms analyzed from 27 haplotypes. Only the polymorphic bases are depicted. The HP2FS haplotype contains a 300-bp segment with derived paralogous gene conversion from HPR (lavender) and a 250-bp region that is highly diverged between subtypes (green/pink). Each allele of the highly diverged region contains a mix of ancestral and derived alleles. The dashes reflect a 2-bp and a 7-bp indel; the other sites shown are individual SNPs. The sequence data used to create this alignment are available online (GenBank: KT923758–KT923784). (b) The frequency of each HP haplotype in four populations. (c) The earlier model of HP structural evolution (interchromosomal non-homologous recombination) would predict the HP1F SNP haplotype background (haplotype B) upstream of HP2 and the HP1S SNP haplotype (haplotype A) downstream of HP2. Additionally, it would predict Form R of the highly diverged region in HP2-Left. However, neither of these predictions was observed in any of the HP2 alleles in this study. ( d ) Both HP1F and HP1S can be created through simple deletions in HP2FS. The dashed lines indicate deleted sequence, while the dashed boxes indicate the sequence required to create each HP1 haplotype. The deletion model is also consistent with the observed SNP haplotype backgrounds surrounding the CNV.

    Journal: Nature genetics

    Article Title: Recurring exon deletions in the haptoglobin (HP) gene associate with lower blood cholesterol levels

    doi: 10.1038/ng.3510

    Figure Lengend Snippet: SNP haplotypes and sequence differences between HP subtypes inform structural history (a) This alignment shows base pair differences between HP structural forms analyzed from 27 haplotypes. Only the polymorphic bases are depicted. The HP2FS haplotype contains a 300-bp segment with derived paralogous gene conversion from HPR (lavender) and a 250-bp region that is highly diverged between subtypes (green/pink). Each allele of the highly diverged region contains a mix of ancestral and derived alleles. The dashes reflect a 2-bp and a 7-bp indel; the other sites shown are individual SNPs. The sequence data used to create this alignment are available online (GenBank: KT923758–KT923784). (b) The frequency of each HP haplotype in four populations. (c) The earlier model of HP structural evolution (interchromosomal non-homologous recombination) would predict the HP1F SNP haplotype background (haplotype B) upstream of HP2 and the HP1S SNP haplotype (haplotype A) downstream of HP2. Additionally, it would predict Form R of the highly diverged region in HP2-Left. However, neither of these predictions was observed in any of the HP2 alleles in this study. ( d ) Both HP1F and HP1S can be created through simple deletions in HP2FS. The dashed lines indicate deleted sequence, while the dashed boxes indicate the sequence required to create each HP1 haplotype. The deletion model is also consistent with the observed SNP haplotype backgrounds surrounding the CNV.

    Article Snippet: Imputation of HP structural variation into cohorts for cholesterol association study A reference panel composed of encoded HP structural alleles and SNPs surrounding the CNV region from the Illumina OMNI, Illumina 1M, and Affymetrix 6.0 arrays was developed and used to impute HP structural variation into cohorts with cholesterol information using Beagle (v2.3.1) imputation software.

    Techniques: Sequencing, Derivative Assay, Homologous Recombination

    Lone HP1S structural alleles segregate on common HP2FS SNP haplotypes SNP haplotype data is shown for three European populations (CEU, IBS, TSI) and one African population (YRI) totaling to 528 haplotypes. SNPs on the left half of the plot exist to the left of the HP duplication (proximal to the centromere), whereas SNPs on the right half of the plot physically reside to the right of the duplication (distal to the centromere). Branch points represent markers at which the depicted haplotypes diverge due to mutation and/or recombination with other haplotypes. The structures are represented on the leaves in order to clarify their relationships to SNP haplotypes, but the CNV and the paralogous gene conversion physically reside within the gap at center of the plot. The African individuals are identified with a dot after the leaf. Arrows with numbers indicate HP1 alleles segregating with the standard HP2 SNP haplotypes for at least 20 kb on both sides of the CNV. The + identifies the SNP haplotype branch which carries HP2FS in almost all sampled Africans, but HP1S in all sampled Europeans. This SNP haplotype is identical downstream of the CNV and differs by a single nucleotide upstream of the CNV. The X indicates the single haplotype observed in this study with apparent recombination in the CNV region (B/A in Figure 2 ). This recombination event appears to be recent because it is identical to standard haplotypes for at least 20 kb on either side of the CNV.

    Journal: Nature genetics

    Article Title: Recurring exon deletions in the haptoglobin (HP) gene associate with lower blood cholesterol levels

    doi: 10.1038/ng.3510

    Figure Lengend Snippet: Lone HP1S structural alleles segregate on common HP2FS SNP haplotypes SNP haplotype data is shown for three European populations (CEU, IBS, TSI) and one African population (YRI) totaling to 528 haplotypes. SNPs on the left half of the plot exist to the left of the HP duplication (proximal to the centromere), whereas SNPs on the right half of the plot physically reside to the right of the duplication (distal to the centromere). Branch points represent markers at which the depicted haplotypes diverge due to mutation and/or recombination with other haplotypes. The structures are represented on the leaves in order to clarify their relationships to SNP haplotypes, but the CNV and the paralogous gene conversion physically reside within the gap at center of the plot. The African individuals are identified with a dot after the leaf. Arrows with numbers indicate HP1 alleles segregating with the standard HP2 SNP haplotypes for at least 20 kb on both sides of the CNV. The + identifies the SNP haplotype branch which carries HP2FS in almost all sampled Africans, but HP1S in all sampled Europeans. This SNP haplotype is identical downstream of the CNV and differs by a single nucleotide upstream of the CNV. The X indicates the single haplotype observed in this study with apparent recombination in the CNV region (B/A in Figure 2 ). This recombination event appears to be recent because it is identical to standard haplotypes for at least 20 kb on either side of the CNV.

    Article Snippet: Imputation of HP structural variation into cohorts for cholesterol association study A reference panel composed of encoded HP structural alleles and SNPs surrounding the CNV region from the Illumina OMNI, Illumina 1M, and Affymetrix 6.0 arrays was developed and used to impute HP structural variation into cohorts with cholesterol information using Beagle (v2.3.1) imputation software.

    Techniques: Mutagenesis

    Alignment of isolated strain 16S rRNA V4 region and representative sequence of Lactobacillus in metagenomics.

    Journal: Toxicological Sciences

    Article Title: Combination of Metagenomics and Culture-Based Methods to Study the Interaction Between Ochratoxin A and Gut Microbiota

    doi: 10.1093/toxsci/kfu128

    Figure Lengend Snippet: Alignment of isolated strain 16S rRNA V4 region and representative sequence of Lactobacillus in metagenomics.

    Article Snippet: The V4 region of the 16S rRNA was amplified by PCR and sequenced by using a MiSeq platform (Illumina, San Diego) according to the manufacturer's instructions.

    Techniques: Isolation, Sequencing

    Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of Aβ and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice

    Journal: Molecular Neurodegeneration

    Article Title: BACE1 elevation engendered by GGA3 deletion increases β-amyloid pathology in association with APP elevation and decreased CHL1 processing in 5XFAD mice

    doi: 10.1186/s13024-018-0239-7

    Figure Lengend Snippet: Schematic diagram of BACE1-mediated processing of CHL1 and APP in old mice. a BACE1 cleaves both APP and CHL1, resulting in the production of Aβ and N-terminal fragment of CHL1 (CHL1_βNTF), respectively, in GGA3WT mice. b Levels of BACE1 are increased in GGA3KO mice, resulting in enhanced processing of CHL1 but not APP. c Levels of transgenic APP increase significantly in GGA3WT;5XFAD mice, resulting in elevated production of Aβ. Conversely, CHL1_βNTF/CHL1_FL ratio is decreased because CHL1_FL is outcompeted by APP for BACE1 cleavage and thus stabilized in GGA3WT;5XFAD mice. d In GGA3KO;5XFAD mice, increased levels of transgenic APP are associated with elevation of BACE1 due to GGA3 deletion, furthering the accumulation of Aβ and the suppression of CHL1_FL cleavage observed in GGA3WT;5XFAD mice

    Article Snippet: The membrane was then incubated in the following antibodies: rabbit monoclonal anti-BACE1 (1:1500; D10E5; Cell signaling); rabbit polyclonal anti-GGA3 (1:1500; 4167; Cell signaling); rabbit polyclonal anti-GGA1 (1:2000; H-215; Santacruz); rabbit polyclonal anti-C-terminal APP antibody to detect mouse and human APP (1:5000; A8717; Sigma Aldrich); goat polyclonal anti-N-terminal CHL1 antibody to detect mouse CHL1_βNTF and CHL1_FL (1:1000; AF2147; R & D Systems); rat monoclonal anti-N-terminal CHL1 antibody to detect human CHL1_βNTF and CHL1_FL (1:2000; MAB2126; R & D Systems); mouse monoclonal anti-human APP antibody which recognizes the amino acid sequence 1–16 in Aβ region (1:10,000; 6E10; Biolegend); mouse monoclonal anti-calnexin to detect mouse and human calnexin (1:2000; 610523; BD biosciences); mouse monoclonal anti-GAPDH (1:10,000; MAP374; Millipore); and mouse monoclonal anti-β-tubulin (1:10,000; JDR.3BR; Sigma-Aldrich).

    Techniques: Mouse Assay, Transgenic Assay

    V4 region and 5′ and 3′ PCR primers. The target nucleotides for the PCR primers are depicted in grey (5′region) and black (3′region). The 5′ and 3′ primers include adaptor Illumina sequences for NexGen sequences

    Journal: Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]

    Article Title: Getting Started with Microbiome Analysis: Sample Acquisition to Bioinformatics

    doi: 10.1002/0471142905.hg1808s82

    Figure Lengend Snippet: V4 region and 5′ and 3′ PCR primers. The target nucleotides for the PCR primers are depicted in grey (5′region) and black (3′region). The 5′ and 3′ primers include adaptor Illumina sequences for NexGen sequences

    Article Snippet: Degenerate PCR primers specific for the V4 region (Eurofins – mwg/operon ( ).

    Techniques: Polymerase Chain Reaction

    The 16 S, 23S and 5S rRNA gene. The expanded 16S gene depicts the variable regions. The target V4 region for microbiome analysis is depicted in black.

    Journal: Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]

    Article Title: Getting Started with Microbiome Analysis: Sample Acquisition to Bioinformatics

    doi: 10.1002/0471142905.hg1808s82

    Figure Lengend Snippet: The 16 S, 23S and 5S rRNA gene. The expanded 16S gene depicts the variable regions. The target V4 region for microbiome analysis is depicted in black.

    Article Snippet: Degenerate PCR primers specific for the V4 region (Eurofins – mwg/operon ( ).

    Techniques: