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  • 94
    Zymo Research cpg dna methylase
    DNMT3a recruitment to the CRE site (−111/−104) is decreased in SLE T cells. A , ChIP was performed using total T cells from four matched pairs of SLE patients and healthy controls ( CON ) and anti-DNMT3a antibody. Immunoprecipitated <t>DNA</t> was analyzed by real-time qPCR using primers detecting a portion of the IL17A promoter harboring the novel CRE site (−111/−104). Ratios between anti-DNMT3a immunoprecipitated and input DNA are shown. Dotted lines associate data from the matched control/SLE pairs. Horizontal bars represent the mean. B , percentage of anti-DNMT3a immunoprecipitated DNA in T cells from the control individual was set to 100%, and relative DNMT3a binding in the corresponding SLE patient was calculated. Values are given as mean ± S.D. ( error bars ). C , Jurkat T cells were transfected with pcDNA3 empty vector ( EV ) or an expression plasmid for DNMT3a. Relative IL-17A mRNA expression was analyzed 5 h after transfection. Values are given as mean ± S.D. from three experiments. D , schematic of <t>CpG-DNA</t> methylation sites within the proximal 195 bp of the IL17A reporter construct. E , pGL3-Basic and IL17A p(−195)-luc reporter plasmids were methylated as outlined under “Experimental Procedures.” Promoter activities of the unmethylated and the methylated reporters were assessed in primary human T cells. Values are given as mean ± S.D. from three experiments.
    Cpg Dna Methylase, supplied by Zymo Research, used in various techniques. Bioz Stars score: 94/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs cpg
    Representative results of the quantitations of the methylation levels by Multiplex-BSP-seq for health control (HC), chronic hepatitis B infection(CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) plasma <t>DNA</t> samples. For each gene, the heat map of the methylation patterns for each <t>CpG</t> is shown. The methylation level (%) measured at each individual CpG site is expressed by the percentage of methylated CpG versus unmethylated CpG sites. HC, CHB, LC and HCC are represented by colored areas of blue, green, violet and red, respectively. The colored area is defined by 25%/75% quantiles. The comparison of the CpG methylation levels for each stage is colored blue to red, in small squares, to indicate the different P values. Representative heat map and methylation plot analysis of 5 target genes; consistently low levels of methylation of all CpGs were observed in GAPDH and steady high levels of CpG(CG1,2 and 7) methylation in KCNV1 are independent of the HCC developmental stage, while the methylation statuses of the other 3 genes (ZNF300, SLC22A20 and SHISA7) varied according to the developmental stage.
    Cpg, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 202 article reviews
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    85
    Thermo Fisher cpg site cg07747299
    Representative results of the quantitations of the methylation levels by Multiplex-BSP-seq for health control (HC), chronic hepatitis B infection(CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) plasma <t>DNA</t> samples. For each gene, the heat map of the methylation patterns for each <t>CpG</t> is shown. The methylation level (%) measured at each individual CpG site is expressed by the percentage of methylated CpG versus unmethylated CpG sites. HC, CHB, LC and HCC are represented by colored areas of blue, green, violet and red, respectively. The colored area is defined by 25%/75% quantiles. The comparison of the CpG methylation levels for each stage is colored blue to red, in small squares, to indicate the different P values. Representative heat map and methylation plot analysis of 5 target genes; consistently low levels of methylation of all CpGs were observed in GAPDH and steady high levels of CpG(CG1,2 and 7) methylation in KCNV1 are independent of the HCC developmental stage, while the methylation statuses of the other 3 genes (ZNF300, SLC22A20 and SHISA7) varied according to the developmental stage.
    Cpg Site Cg07747299, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Illumina Inc cpgs
    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT <t>CpGs</t> are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative <t>DNA</t> methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p
    Cpgs, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 94/100, based on 1696 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Active Motif cpgs
    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT <t>CpGs</t> are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative <t>DNA</t> methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p
    Cpgs, supplied by Active Motif, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 7 article reviews
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    85
    InvivoGen cpg oligodinucleotide 1826 cpg
    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT <t>CpGs</t> are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative <t>DNA</t> methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p
    Cpg Oligodinucleotide 1826 Cpg, supplied by InvivoGen, 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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    Average 85 stars, based on 8 article reviews
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    95
    Millipore 3 tamra cpg
    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT <t>CpGs</t> are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative <t>DNA</t> methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p
    3 Tamra Cpg, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Coley Pharmaceutical cpg odn 1826 cpg
    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT <t>CpGs</t> are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative <t>DNA</t> methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p
    Cpg Odn 1826 Cpg, supplied by Coley Pharmaceutical, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Bioneer Corporation cpg odns cpg odns
    The relative mRNA expressions of Mx (a); IgM (b); CC (c); and interleukin 1- β (d) in cobia intestine after stimulating with different <t>CpG</t> <t>ODNs</t> (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values are represented as “fold change” and compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different letters are significantly different ( P ≤ 0.05).
    Cpg Odns Cpg Odns, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    TIB MOLBIOL cpg 2006 oligonucleotide cpg
    The relative mRNA expressions of Mx (a); IgM (b); CC (c); and interleukin 1- β (d) in cobia intestine after stimulating with different <t>CpG</t> <t>ODNs</t> (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values are represented as “fold change” and compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different letters are significantly different ( P ≤ 0.05).
    Cpg 2006 Oligonucleotide Cpg, supplied by TIB MOLBIOL, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Dynavax Technologies cpg a
    The relative mRNA expressions of Mx (a); IgM (b); CC (c); and interleukin 1- β (d) in cobia intestine after stimulating with different <t>CpG</t> <t>ODNs</t> (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values are represented as “fold change” and compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different letters are significantly different ( P ≤ 0.05).
    Cpg A, supplied by Dynavax Technologies, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    InvivoGen cpg a
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Cpg A, supplied by InvivoGen, used in various techniques. Bioz Stars score: 91/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Illumina Inc 1505 cpgs
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    1505 Cpgs, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Qiagen cpg sites
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Cpg Sites, supplied by Qiagen, used in various techniques. Bioz Stars score: 94/100, based on 1355 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    InvivoGen oligodeoxynucleotides cpg 1585 cpg a
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Oligodeoxynucleotides Cpg 1585 Cpg A, supplied by InvivoGen, 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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    85
    InvivoGen cpg motifs cpg odn
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Cpg Motifs Cpg Odn, supplied by InvivoGen, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Oligos Etc cpg oligodeoxynucleotides 1826 cpg
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Cpg Oligodeoxynucleotides 1826 Cpg, supplied by Oligos Etc, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Enzo Biochem cpg a
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
    Cpg A, supplied by Enzo Biochem, 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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    GeneWorks odn1826 cpgs
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
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    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
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    Qiagen cpg mlh1
    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
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    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with <t>CpG-A.</t> ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P
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    Thermo Fisher cpg
    TLR activation by OLA. TLR2- (A), TLR4- (B) and TLR9- (C) transfected human embryonic kidney cells (HEK293) cells were incubated with OLA at three different concentrations and the respective positive controls (Pam3Cys, LPS and <t>CpG)</t> for 24h. Activation was assessed by measuring human (h)IL-8 production by ELISA. Results represent mean values ± SEM from one representative experiment.
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    Image Search Results


    DNMT3a recruitment to the CRE site (−111/−104) is decreased in SLE T cells. A , ChIP was performed using total T cells from four matched pairs of SLE patients and healthy controls ( CON ) and anti-DNMT3a antibody. Immunoprecipitated DNA was analyzed by real-time qPCR using primers detecting a portion of the IL17A promoter harboring the novel CRE site (−111/−104). Ratios between anti-DNMT3a immunoprecipitated and input DNA are shown. Dotted lines associate data from the matched control/SLE pairs. Horizontal bars represent the mean. B , percentage of anti-DNMT3a immunoprecipitated DNA in T cells from the control individual was set to 100%, and relative DNMT3a binding in the corresponding SLE patient was calculated. Values are given as mean ± S.D. ( error bars ). C , Jurkat T cells were transfected with pcDNA3 empty vector ( EV ) or an expression plasmid for DNMT3a. Relative IL-17A mRNA expression was analyzed 5 h after transfection. Values are given as mean ± S.D. from three experiments. D , schematic of CpG-DNA methylation sites within the proximal 195 bp of the IL17A reporter construct. E , pGL3-Basic and IL17A p(−195)-luc reporter plasmids were methylated as outlined under “Experimental Procedures.” Promoter activities of the unmethylated and the methylated reporters were assessed in primary human T cells. Values are given as mean ± S.D. from three experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: cAMP-responsive Element Modulator (CREM)? Protein Induces Interleukin 17A Expression and Mediates Epigenetic Alterations at the Interleukin-17A Gene Locus in Patients with Systemic Lupus Erythematosus *

    doi: 10.1074/jbc.M111.299313

    Figure Lengend Snippet: DNMT3a recruitment to the CRE site (−111/−104) is decreased in SLE T cells. A , ChIP was performed using total T cells from four matched pairs of SLE patients and healthy controls ( CON ) and anti-DNMT3a antibody. Immunoprecipitated DNA was analyzed by real-time qPCR using primers detecting a portion of the IL17A promoter harboring the novel CRE site (−111/−104). Ratios between anti-DNMT3a immunoprecipitated and input DNA are shown. Dotted lines associate data from the matched control/SLE pairs. Horizontal bars represent the mean. B , percentage of anti-DNMT3a immunoprecipitated DNA in T cells from the control individual was set to 100%, and relative DNMT3a binding in the corresponding SLE patient was calculated. Values are given as mean ± S.D. ( error bars ). C , Jurkat T cells were transfected with pcDNA3 empty vector ( EV ) or an expression plasmid for DNMT3a. Relative IL-17A mRNA expression was analyzed 5 h after transfection. Values are given as mean ± S.D. from three experiments. D , schematic of CpG-DNA methylation sites within the proximal 195 bp of the IL17A reporter construct. E , pGL3-Basic and IL17A p(−195)-luc reporter plasmids were methylated as outlined under “Experimental Procedures.” Promoter activities of the unmethylated and the methylated reporters were assessed in primary human T cells. Values are given as mean ± S.D. from three experiments.

    Article Snippet: To investigate the effects of CpG-DNA methylation on IL17A promoter activity, we methylated both the 195-bp IL17A reporter construct and the empty pGL3 plasmid, using CpG-DNA methylase (Zymo Research) according to the manufacturer's instructions.

    Techniques: Chromatin Immunoprecipitation, Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Transfection, Plasmid Preparation, Expressing, DNA Methylation Assay, Construct, Methylation

    Decreased CpG-DNA methylation in IL-17A-secreting and SLE T cells. A , CpG sites within the CNS and the proximal promoter ( PP ) of the human IL17A gene are indicated. B , naïve CD4 + T cells from healthy blood donors that had been stimulated with anti-CD3/anti-CD28 antibodies for 72 h followed by stimulation with PMA/ionomycin for another 5 h were subjected to an IL-17A secretion assay. ChIP analyses were performed in both IL-17A-enriched T cells ( black bars ) and non-IL-17A-secreting T cells ( gray bars ), using an antibody that specifically detects methylated CpG sequences. Methylated DNA was recovered, and CNS and proximal promoter regions were amplified by real-time qPCR. Completely methylated (input, 100%) and unmethylated human DNA samples (negative control, 0%) were included. Values are given as mean ± S.D. ( error bars ) from four independent experiments. C , total T cells from six individually matched SLE ( gray bars ) and healthy control individuals ( CON ; black bars ) were subjected to CpG-DNA immunoprecipitation. The percentage of methylated DNA is given as mean ± S.D.

    Journal: The Journal of Biological Chemistry

    Article Title: cAMP-responsive Element Modulator (CREM)? Protein Induces Interleukin 17A Expression and Mediates Epigenetic Alterations at the Interleukin-17A Gene Locus in Patients with Systemic Lupus Erythematosus *

    doi: 10.1074/jbc.M111.299313

    Figure Lengend Snippet: Decreased CpG-DNA methylation in IL-17A-secreting and SLE T cells. A , CpG sites within the CNS and the proximal promoter ( PP ) of the human IL17A gene are indicated. B , naïve CD4 + T cells from healthy blood donors that had been stimulated with anti-CD3/anti-CD28 antibodies for 72 h followed by stimulation with PMA/ionomycin for another 5 h were subjected to an IL-17A secretion assay. ChIP analyses were performed in both IL-17A-enriched T cells ( black bars ) and non-IL-17A-secreting T cells ( gray bars ), using an antibody that specifically detects methylated CpG sequences. Methylated DNA was recovered, and CNS and proximal promoter regions were amplified by real-time qPCR. Completely methylated (input, 100%) and unmethylated human DNA samples (negative control, 0%) were included. Values are given as mean ± S.D. ( error bars ) from four independent experiments. C , total T cells from six individually matched SLE ( gray bars ) and healthy control individuals ( CON ; black bars ) were subjected to CpG-DNA immunoprecipitation. The percentage of methylated DNA is given as mean ± S.D.

    Article Snippet: To investigate the effects of CpG-DNA methylation on IL17A promoter activity, we methylated both the 195-bp IL17A reporter construct and the empty pGL3 plasmid, using CpG-DNA methylase (Zymo Research) according to the manufacturer's instructions.

    Techniques: DNA Methylation Assay, Chromatin Immunoprecipitation, Methylation, Amplification, Real-time Polymerase Chain Reaction, Negative Control, Immunoprecipitation

    Representative results of the quantitations of the methylation levels by Multiplex-BSP-seq for health control (HC), chronic hepatitis B infection(CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) plasma DNA samples. For each gene, the heat map of the methylation patterns for each CpG is shown. The methylation level (%) measured at each individual CpG site is expressed by the percentage of methylated CpG versus unmethylated CpG sites. HC, CHB, LC and HCC are represented by colored areas of blue, green, violet and red, respectively. The colored area is defined by 25%/75% quantiles. The comparison of the CpG methylation levels for each stage is colored blue to red, in small squares, to indicate the different P values. Representative heat map and methylation plot analysis of 5 target genes; consistently low levels of methylation of all CpGs were observed in GAPDH and steady high levels of CpG(CG1,2 and 7) methylation in KCNV1 are independent of the HCC developmental stage, while the methylation statuses of the other 3 genes (ZNF300, SLC22A20 and SHISA7) varied according to the developmental stage.

    Journal: Clinical Epigenetics

    Article Title: Genome-wide methylation profiling of the different stages of hepatitis B virus-related hepatocellular carcinoma development in plasma cell-free DNA reveals potential biomarkers for early detection and high-risk monitoring of hepatocellular carcinoma

    doi: 10.1186/1868-7083-6-30

    Figure Lengend Snippet: Representative results of the quantitations of the methylation levels by Multiplex-BSP-seq for health control (HC), chronic hepatitis B infection(CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) plasma DNA samples. For each gene, the heat map of the methylation patterns for each CpG is shown. The methylation level (%) measured at each individual CpG site is expressed by the percentage of methylated CpG versus unmethylated CpG sites. HC, CHB, LC and HCC are represented by colored areas of blue, green, violet and red, respectively. The colored area is defined by 25%/75% quantiles. The comparison of the CpG methylation levels for each stage is colored blue to red, in small squares, to indicate the different P values. Representative heat map and methylation plot analysis of 5 target genes; consistently low levels of methylation of all CpGs were observed in GAPDH and steady high levels of CpG(CG1,2 and 7) methylation in KCNV1 are independent of the HCC developmental stage, while the methylation statuses of the other 3 genes (ZNF300, SLC22A20 and SHISA7) varied according to the developmental stage.

    Article Snippet: The in vitro methylated DNA from the HepG2 cells was obtained using the CpG (M. SssI) methyltransferase (NEB, MA,USA) treatment and used as a positive control.

    Techniques: Methylation, Multiplex Assay, Infection, CpG Methylation Assay

    Receiver operating characteristics (ROC) and multiple univariate logistic regression analyses for using CpGs to distinguish between hepatocellular carcinoma (HCC) developmental stages are shown according to the CpG position and disease stage (HC + CHB versus LC + HCC or HC + CHB + LC versus HCC). (A) Receiver operating characteristic (ROC) curves for ZNF300, SLC22A20 and SHISA7. Complete DNA methylation data from all four stages of HCC development were used to construct the ROC curves. The ROC curves plot the sensitivity versus 100-specificity. Upper panel: a lower cut-off value was used to distinguish between (LC + HCC)/ (HC + CHB).Lower panel: a higher cut-off value was used to distinguish between HCC/(HC + CHB + LC). (B) A multiple univariate logistic regression analysis was performed using the CpG methylation patterns to evaluate the association between gene methylation and the stage of HCC development. Relationship between the CpG methylation (odds ratios) and the developmental stage. To separate (LC + HCC)/ (HC + CHB) and HCC/(HC + CHB + LC), both univariate (which considers the methylation levels) and multivariate (which also considers the age and gender) logistic regressions were performed using CpG methylation data for ZNF300, SLC22A20 and SHISA7.

    Journal: Clinical Epigenetics

    Article Title: Genome-wide methylation profiling of the different stages of hepatitis B virus-related hepatocellular carcinoma development in plasma cell-free DNA reveals potential biomarkers for early detection and high-risk monitoring of hepatocellular carcinoma

    doi: 10.1186/1868-7083-6-30

    Figure Lengend Snippet: Receiver operating characteristics (ROC) and multiple univariate logistic regression analyses for using CpGs to distinguish between hepatocellular carcinoma (HCC) developmental stages are shown according to the CpG position and disease stage (HC + CHB versus LC + HCC or HC + CHB + LC versus HCC). (A) Receiver operating characteristic (ROC) curves for ZNF300, SLC22A20 and SHISA7. Complete DNA methylation data from all four stages of HCC development were used to construct the ROC curves. The ROC curves plot the sensitivity versus 100-specificity. Upper panel: a lower cut-off value was used to distinguish between (LC + HCC)/ (HC + CHB).Lower panel: a higher cut-off value was used to distinguish between HCC/(HC + CHB + LC). (B) A multiple univariate logistic regression analysis was performed using the CpG methylation patterns to evaluate the association between gene methylation and the stage of HCC development. Relationship between the CpG methylation (odds ratios) and the developmental stage. To separate (LC + HCC)/ (HC + CHB) and HCC/(HC + CHB + LC), both univariate (which considers the methylation levels) and multivariate (which also considers the age and gender) logistic regressions were performed using CpG methylation data for ZNF300, SLC22A20 and SHISA7.

    Article Snippet: The in vitro methylated DNA from the HepG2 cells was obtained using the CpG (M. SssI) methyltransferase (NEB, MA,USA) treatment and used as a positive control.

    Techniques: DNA Methylation Assay, Construct, CpG Methylation Assay, Methylation

    Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT CpGs are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative DNA methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p

    Journal: PLoS Medicine

    Article Title: Role of DNA Methylation and Epigenetic Silencing of HAND2 in Endometrial Cancer Development

    doi: 10.1371/journal.pmed.1001551

    Figure Lengend Snippet: Discovery of HAND2 methylation as a core feature in endometrial cancer. (A) Volcano plot of epigenome-wide differential DNAme analysis for all 27,578 probes. The x -axis indicates the median β-value difference between the normal and cancerous endometrial samples (median[cancer] – median[normal]), while the y -axis indicates the −log e scale of q -values obtained from a supervised logistic regression analysis testing the association of methylation with normal/cancer status (Set 1). Stem cell PCGT CpGs are highlighted in green, the two HAND2 CpGs in red. 353 PCGT CpGs are hypermethylated, and 19 PCGT CpGs are hypomethylated, with enrichment odds ratio (OR) and p -value (P) obtained from a one-sided Fisher's exact test. The horizontal dotted lines mark the significance cutoffs. (B) Integrative DNA methylome (DNAm)–interactome analysis to identify differential methylation hotspots in the network. Briefly, edge weights in the interactome network reflect the combined differential methylation statistics (absolute values) of the genes making up the edge (the CpG closest to the transcription start site [TSS] of the gene was chosen). A spin-glass module detection algorithm was subsequently used to identify subnetworks where the average edge weight (“modularity”) is higher than random, as assessed by randomly rewiring the network preserving node degrees. Statistical significance of the subnetworks was further assessed by comparing their modularities to those obtained by permuting differential methylation statistics over the network. Subnetworks with p

    Article Snippet: Analyses Synopsis We analysed the DNA methylation (DNAme) of ∼27,000 CpGs (Illumina Infinium HumanMethylation27K BeadChip) in normal and cancerous endometrial tissue (Set 1) and applied a novel integrative epigenome-transcriptome-interactome approach (by also adding data from Set 2) to identify epigenetically deregulated interactome hotspots of functional significance associated with the phenotype of interest, i.e., endometrial cancer.

    Techniques: Methylation, Preserving

    The relative mRNA expressions of Mx (a); IgM (b); CC (c); and interleukin 1- β (d) in cobia intestine after stimulating with different CpG ODNs (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values are represented as “fold change” and compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different letters are significantly different ( P ≤ 0.05).

    Journal: Journal of Immunology Research

    Article Title: The Effect of TLR9 Agonist CpG Oligodeoxynucleotides on the Intestinal Immune Response of Cobia (Rachycentron canadum)

    doi: 10.1155/2014/273284

    Figure Lengend Snippet: The relative mRNA expressions of Mx (a); IgM (b); CC (c); and interleukin 1- β (d) in cobia intestine after stimulating with different CpG ODNs (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values are represented as “fold change” and compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different letters are significantly different ( P ≤ 0.05).

    Article Snippet: CpG ODNs CpG ODNs were purchased from Bioneer (Korea).

    Techniques: Polymerase Chain Reaction, Expressing

    The relative mRNA expressions of TLR 9A (a); TLR 9B (b); and MyD88 (c) in cobia intestine after stimulating with different CpG ODNs (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values represented as “fold change” were compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different alphabetical letters are significantly different ( P ≤ 0.05).

    Journal: Journal of Immunology Research

    Article Title: The Effect of TLR9 Agonist CpG Oligodeoxynucleotides on the Intestinal Immune Response of Cobia (Rachycentron canadum)

    doi: 10.1155/2014/273284

    Figure Lengend Snippet: The relative mRNA expressions of TLR 9A (a); TLR 9B (b); and MyD88 (c) in cobia intestine after stimulating with different CpG ODNs (1668, 2395) and ODN 2137 as control measured by quantitative real-tie PCR at 1, 3, 6, and 10 days after stimulation. The expression values represented as “fold change” were compared to the noninjected control samples and β -actin was used as a reference gene. The results are presented as the mean ± SD ( n = 2) and mean values with different alphabetical letters are significantly different ( P ≤ 0.05).

    Article Snippet: CpG ODNs CpG ODNs were purchased from Bioneer (Korea).

    Techniques: Polymerase Chain Reaction, Expressing

    Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A. ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P

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

    Article Title: Enhanced susceptibility to chemically induced colitis caused by excessive endosomal TLR signaling in LRBA-deficient mice

    doi: 10.1073/pnas.1901407116

    Figure Lengend Snippet: Enhanced PI3K/AKT/mTOR signaling in Lrba −/− BMDCs and BMpDCs. ( A ) Immunoblot analysis of FOXO1 and FOXO3 in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A. ( B and C ) Immunoblot analysis of phosphorylated and total mTOR, AKT, S6, and 4E-BP1 in Lrba +/+ , Lrba +/− , and Lrba −/− ( B ) BMpDCs and ( C ) BMDCs stimulated with CpG-A for 16 h ( B ) or for the indicated times ( C ). ( D , Left ) Flow cytometry of Lrba +/+ and Lrba −/− splenocytes, assessing expression of p-AKT and p-S6 by CD11c + DCs. ( D , Right ) Quantification of p-AKT and p-S6 mean fluorescence intensity (MFI) on CD11c + DCs. ( E and F ) Concentration of IFN-α in the culture supernatant of ( E ) BMDCs or ( F ) BMpDCs pretreated with PI3K and mTOR inhibitors for 3 h, then stimulated with CpG-A for 16 h ( n = 3 independent cultures of each genotype from separate mice). ( G ) RT-qPCR analysis of the indicated mRNAs in Lrba +/+ and Lrba −/− BMpDCs after stimulation with CpG-A for 16 h. ( H ) Protein array analysis of the culture supernatant from BMpDCs stimulated with CpG-A. Each symbol ( D ) represents an individual mouse. * P

    Article Snippet: Peritoneal m acrophages were stimulated for 16 h with the following TLR ligands: poly (I:C) (200 μg/mL; Invivogen), R848 (20 ng/mL; Enzo Life Sciences), LPS (10 ng/mL; Enzo Life Sciences), CpG-A (100 μg/mL; Invivogen), or CpG-B (200 ng/mL; Invivogen).

    Techniques: Flow Cytometry, Cytometry, Expressing, Fluorescence, Concentration Assay, Mouse Assay, Quantitative RT-PCR, Protein Array

    Enhanced activation of IRF3 and IRF7 in Lrba −/− cells stimulated with TLR3 or TLR9 ligands. ( A ) Immunoblot analysis of phosphorylated (p) or total IKKα/β, NF-κB p65, ERK1/2, and IκBα in Lrba +/+ , Lrba +/− , and Lrba −/− BMDCs after stimulation with CpG-B. ( B and C ) Immunoblot analysis of ( B ) IRF7, NF-κB p65, and ( C ) IRF3 in nuclear and cytoplasmic fractions of lysates of Lrba +/+ and Lrba −/− BMpDCs after stimulation with ( B ) CpG-A and (C) poly(I:C). ( D ) Immunoblot analysis of phosphorylated (p) and total IRF3 in Lrba +/+ , Lrba +/− , and Lrba −/− BMpDCs after stimulation with poly(I:C). Data are representative of three independent experiments.

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

    Article Title: Enhanced susceptibility to chemically induced colitis caused by excessive endosomal TLR signaling in LRBA-deficient mice

    doi: 10.1073/pnas.1901407116

    Figure Lengend Snippet: Enhanced activation of IRF3 and IRF7 in Lrba −/− cells stimulated with TLR3 or TLR9 ligands. ( A ) Immunoblot analysis of phosphorylated (p) or total IKKα/β, NF-κB p65, ERK1/2, and IκBα in Lrba +/+ , Lrba +/− , and Lrba −/− BMDCs after stimulation with CpG-B. ( B and C ) Immunoblot analysis of ( B ) IRF7, NF-κB p65, and ( C ) IRF3 in nuclear and cytoplasmic fractions of lysates of Lrba +/+ and Lrba −/− BMpDCs after stimulation with ( B ) CpG-A and (C) poly(I:C). ( D ) Immunoblot analysis of phosphorylated (p) and total IRF3 in Lrba +/+ , Lrba +/− , and Lrba −/− BMpDCs after stimulation with poly(I:C). Data are representative of three independent experiments.

    Article Snippet: Peritoneal m acrophages were stimulated for 16 h with the following TLR ligands: poly (I:C) (200 μg/mL; Invivogen), R848 (20 ng/mL; Enzo Life Sciences), LPS (10 ng/mL; Enzo Life Sciences), CpG-A (100 μg/mL; Invivogen), or CpG-B (200 ng/mL; Invivogen).

    Techniques: Activation Assay

    Increased type I IFN responses to endosomal TLR stimulation in Lrba −/− DCs. ( A – E ) In vitro differentiated BMDCs or BMpDCs were stimulated with TLR ligands for 16 h. Concentration of ( A ) IFN-β or ( B ) IL-23 in the culture supernatant of BMDCs after stimulation with different TLR ligands ( n = 4 independent cultures of each genotype from separate mice). ( C ) Concentration of IFN-α in the culture supernatant of BMpDCs after stimulation with the indicated ligands ( n = 3 independent cultures of each genotype from separate mice). ( D and E ) Serum concentration of ( D ) IFN-α and ( E ) IL-6 4 h after injection of DOTAP-encapsulated CpG-A into mice ( n = 4 mice per genotype). ( F and G ) BM transplantation was performed using mice of the indicated genotypes (donor > recipient). ( F ) Body weight (% relative to weight on day 0) and ( G ) DAI on day 10 after initiation of DSS treatment. Lrba +/+ Ifnar1 +/+ > Rag2 −/− ( n = 4), Lrba −/− Ifnar1 +/+ > Rag2 −/− ( n = 4), Lrba +/+ Ifnar1 macro−1/macro−1 > Rag2 −/− ( n = 4), and Lrba −/− Ifnar1 macro−1/macro−1 > Rag2 −/− ( n = 4). * P

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

    Article Title: Enhanced susceptibility to chemically induced colitis caused by excessive endosomal TLR signaling in LRBA-deficient mice

    doi: 10.1073/pnas.1901407116

    Figure Lengend Snippet: Increased type I IFN responses to endosomal TLR stimulation in Lrba −/− DCs. ( A – E ) In vitro differentiated BMDCs or BMpDCs were stimulated with TLR ligands for 16 h. Concentration of ( A ) IFN-β or ( B ) IL-23 in the culture supernatant of BMDCs after stimulation with different TLR ligands ( n = 4 independent cultures of each genotype from separate mice). ( C ) Concentration of IFN-α in the culture supernatant of BMpDCs after stimulation with the indicated ligands ( n = 3 independent cultures of each genotype from separate mice). ( D and E ) Serum concentration of ( D ) IFN-α and ( E ) IL-6 4 h after injection of DOTAP-encapsulated CpG-A into mice ( n = 4 mice per genotype). ( F and G ) BM transplantation was performed using mice of the indicated genotypes (donor > recipient). ( F ) Body weight (% relative to weight on day 0) and ( G ) DAI on day 10 after initiation of DSS treatment. Lrba +/+ Ifnar1 +/+ > Rag2 −/− ( n = 4), Lrba −/− Ifnar1 +/+ > Rag2 −/− ( n = 4), Lrba +/+ Ifnar1 macro−1/macro−1 > Rag2 −/− ( n = 4), and Lrba −/− Ifnar1 macro−1/macro−1 > Rag2 −/− ( n = 4). * P

    Article Snippet: Peritoneal m acrophages were stimulated for 16 h with the following TLR ligands: poly (I:C) (200 μg/mL; Invivogen), R848 (20 ng/mL; Enzo Life Sciences), LPS (10 ng/mL; Enzo Life Sciences), CpG-A (100 μg/mL; Invivogen), or CpG-B (200 ng/mL; Invivogen).

    Techniques: In Vitro, Concentration Assay, Mouse Assay, Injection, Transplantation Assay

    TLR activation by OLA. TLR2- (A), TLR4- (B) and TLR9- (C) transfected human embryonic kidney cells (HEK293) cells were incubated with OLA at three different concentrations and the respective positive controls (Pam3Cys, LPS and CpG) for 24h. Activation was assessed by measuring human (h)IL-8 production by ELISA. Results represent mean values ± SEM from one representative experiment.

    Journal: PLoS ONE

    Article Title: Oocyst-Derived Extract of Toxoplasma Gondii Serves as Potent Immunomodulator in a Mouse Model of Birch Pollen Allergy

    doi: 10.1371/journal.pone.0155081

    Figure Lengend Snippet: TLR activation by OLA. TLR2- (A), TLR4- (B) and TLR9- (C) transfected human embryonic kidney cells (HEK293) cells were incubated with OLA at three different concentrations and the respective positive controls (Pam3Cys, LPS and CpG) for 24h. Activation was assessed by measuring human (h)IL-8 production by ELISA. Results represent mean values ± SEM from one representative experiment.

    Article Snippet: Plated HEK293 cells (5 x 105 /ml) were stimulated with OLA/polymyxin B (1, 10 and 100 μg/ml) and the respective controls Pam3Cys (1 μg/ml), LPS (2 ng/ml), CpG (100 μg/ml) or medium/polymyxin B for 24 h. Levels of human IL-8 were then measured in supernatants by ELISA (eBioscience).

    Techniques: Activation Assay, Transfection, Incubation, Enzyme-linked Immunosorbent Assay