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massarray quantitative dna methylation analysis  (Sequenom)

 
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    Structured Review

    Sequenom massarray quantitative dna methylation analysis
    ZNF582-AS1 expression was regulated by <t>DNA</t> <t>methylation</t> in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom <t>MassARRAY</t> quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated
    Massarray Quantitative Dna Methylation Analysis, supplied by Sequenom, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/massarray+quantitative+methylation+analysis/massarray+quantitative+methylation+analysis/pmc07945252-212-54-53
    Average 90 stars, based on 1 article reviews
    massarray quantitative dna methylation analysis - by Bioz Stars, 2026-10
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    Images

    1) Product Images from "Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1"

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-021-01889-8

    ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated
    Figure Legend Snippet: ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Techniques Used: Expressing, DNA Methylation Assay, Sequencing, Comparison, Methylation

    Related Articles

    Methylation:

    Article Title: Gender dimorphism in hepatocarcinogenesis—DNA methylation modification regulated X‐chromosome inactivation escape molecule XIST
    Article Snippet: .. Methylation of CpG sites of the Xist first exon region in liver tissues between 10 WT and 10 Tet2 −/− female mice was quantified by Sequenom MassARRAY methylation profiling. ..

    Article Title: Gender dimorphism in hepatocarcinogenesis—DNA methylation modification regulated X‐chromosome inactivation escape molecule XIST
    Article Snippet: .. Methylation of XIST was detected by Sequenom MassARRAY methylation profiling between HCC tissues (T) and adjacent normal liver tissues (L). ..

    Article Title: Exposure to Juvenile Stress Induces Epigenetic Alterations in the GABAergic System in Rats.
    Article Snippet: .. Sequenom Epityper MassARRAY Methylation and Spectra Analysis Samples were desalted and spotted on a 384-alternative patch SpectroCHIP (Se- quenom, San Diego, CA, USA) using the MassARRAY nanodispenser, and spectra were analyzed by the MassARRAY Analyzer Compact matrix-assisted laser desorption/ioniza- tion–time of flight (MALDI-TOF MS) (Sequenom, San Diego, CA, USA). .. Spectra were elaborated by the Epityper 1.2 software (Sequenom, San Diego, CA, USA), which provides methylation values for each CpG unit as a percentage.

    Article Title: DNA Hyper-methylation Associated With Schizophrenia May Lead to Increased Levels of Autoantibodies
    Article Snippet: .. A Sequenom MassARRAY quantitative methylation assay was performed to measure the methylation levels of candidate DMS cg14341177 and cg13978347 in the discovery cohort. .. There are 4 CpG sites on cg14341177, in which the CpG2 site showed the most significant changes in methylation levels and seemed as the leading CpG site.

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..

    other:

    Article Title: Prediction of treatment response to antipsychotic drugs for precision medicine approach to schizophrenia: randomized trials and multiomics analysis
    Article Snippet: The methylation profiling was validated by two methods including Illumina-sequencing-based BSP and Sequenom MassARRAY® Methylation.

    Article Title: Prediction of treatment response to antipsychotic drugs for precision medicine approach to schizophrenia: randomized trials and multiomics analysis.
    Article Snippet: For participants with methylation data, technical replication (see Technical replication of DNA methylation profiling in Additional file 1) was conducted: 194 participants were randomly chosen for Illumina sequencing-based BSP detection to verify the chip detection site results, and among these 194 participants, 20 were further randomly selected to undergo Sequenom MassARRAY® Methylation validation.

    Expressing:

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..

    Western Blot:

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..

    Real-time Polymerase Chain Reaction:

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..

    Agarose Gel Electrophoresis:

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..

    Methylated DNA Immunoprecipitation:

    Article Title: Gender dimorphism in hepatocarcinogenesis-DNA methylation modification regulated X-chromosome inactivation escape molecule XIST.
    Article Snippet: .. Scale bar, 50 μm. (G and H) The expression of YY1 in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by western blot analysis. (I) Expression of YY1 and XIST in HCC-1016 cells (knocked down YY1 in HCC-1016 cells by transfecting siRNA) by qPCR. (J) XIST was quantified using DNA agarose gel electrophoresis. (K) The 5-methylcytosine (5mC) level in the XIST promoter region by MeDIP assay and qPCR. was quantified by Sequenom MassARRAY methylation profiling. ..



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    During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through <t>methylation-dependent</t> transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom <t>MassARRAY</t> assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).
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    Image Search Results


    Comparison of methylation data between MeDIP/NimbleGen Promoter + CpGi Array and Sequenom MassArray: Stk31 CpGi promoter region . The Stk31 gene region with a CpGi promoter region that is methylated in the liver, but not in ES or testis cells. The box indicates the position of the methylation peak in the liver that overlaps with the CpGi promoter region analyzed by Sequenom. In the epigram of Sequenom methylation analysis shown at the bottom panel, the blue circles indicate 100% methylation, green circles indicate around 50%methylation and yellow circles indicate 0% methylation. The results from biological duplicate samples are shown. The bottom lane (Sequenom Methylation Analysis) shows the results of analysis of testis DNA after in vitro methylation using M. Sss1 methylase (New England Biolabs).

    Journal: BMC Genomics

    Article Title: Genome-wide survey reveals dynamic widespread tissue-specific changes in DNA methylation during development

    doi: 10.1186/1471-2164-12-231

    Figure Lengend Snippet: Comparison of methylation data between MeDIP/NimbleGen Promoter + CpGi Array and Sequenom MassArray: Stk31 CpGi promoter region . The Stk31 gene region with a CpGi promoter region that is methylated in the liver, but not in ES or testis cells. The box indicates the position of the methylation peak in the liver that overlaps with the CpGi promoter region analyzed by Sequenom. In the epigram of Sequenom methylation analysis shown at the bottom panel, the blue circles indicate 100% methylation, green circles indicate around 50%methylation and yellow circles indicate 0% methylation. The results from biological duplicate samples are shown. The bottom lane (Sequenom Methylation Analysis) shows the results of analysis of testis DNA after in vitro methylation using M. Sss1 methylase (New England Biolabs).

    Article Snippet: Sequenom MassArray quantitative methylation analysis of bisulfite treated DNA was performed as an additional means to confirm DNA methylation peaks obtained by MeDIP/NimbleGen Array for both selected and randomly chosen MeDIP methylated regions.

    Techniques: Methylation, Methylated DNA Immunoprecipitation, In Vitro

    ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Article Snippet: TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells.

    Techniques: Expressing, DNA Methylation Assay, Sequencing, Comparison, Methylation

    DNA methylation analysis of chicken KLF7 in adipose tissues. a . The CpG density of the genomic region of chicken KLF7 analyzed by CpGplot (Version 6.6.0). b . The CpG loci analyzed in the promoter and Exon 2. c . The levels of DNA methylation of CpG loci analyzed. d . The comparison of DNA methylation levels between the promoter and Exon 2. Asterisks indicate a significant difference between the promoter and Exon 2 (Student’s t test, *** P < 0.001)

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: DNA methylation analysis of chicken KLF7 in adipose tissues. a . The CpG density of the genomic region of chicken KLF7 analyzed by CpGplot (Version 6.6.0). b . The CpG loci analyzed in the promoter and Exon 2. c . The levels of DNA methylation of CpG loci analyzed. d . The comparison of DNA methylation levels between the promoter and Exon 2. Asterisks indicate a significant difference between the promoter and Exon 2 (Student’s t test, *** P < 0.001)

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay, Comparison

    Datasets characteristics of DNA  methylation  of chicken KLF7 in abdominal adipose tissues

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Datasets characteristics of DNA methylation of chicken KLF7 in abdominal adipose tissues

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Association analysis of DNA methylation with KLF7 transcripts and blood metabolic indicators. a . The r value of the Spearman correlation analysis. b . The P value of the Spearman correlation analysis. c . The number of samples used in the Spearman correlation analysis. d . The regression analysis between the DNA methylation of PCpG6 and KLF7 transcripts in chicken adipose tissue

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Association analysis of DNA methylation with KLF7 transcripts and blood metabolic indicators. a . The r value of the Spearman correlation analysis. b . The P value of the Spearman correlation analysis. c . The number of samples used in the Spearman correlation analysis. d . The regression analysis between the DNA methylation of PCpG6 and KLF7 transcripts in chicken adipose tissue

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Schematic representation of the relationship of DNA methylation, KLF7 transcripts, glycaemia and abdominal fat content in chicken. The negative associations marked red and blue were confirmed by the reference [ , ], respectively

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Schematic representation of the relationship of DNA methylation, KLF7 transcripts, glycaemia and abdominal fat content in chicken. The negative associations marked red and blue were confirmed by the reference [ , ], respectively

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Oligonucleotides used in the current study

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Oligonucleotides used in the current study

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: Methylation

    During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

    Journal: Chinese Journal of Cancer Research

    Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

    doi: 10.21147/j.issn.1000-9604.2019.06.09

    Figure Lengend Snippet: During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

    Article Snippet: The methylation status of Stomatin promoter was detected using Sequenom MassARRAY quantitative methylation analysis platform (BGI tech, Shenzhen, China).

    Techniques: Methylation, Real-time Polymerase Chain Reaction, Microarray, Sequenom Massarray Assay, Two Tailed Test, Expressing, Western Blot, Control