tissue dna quantitative methylation specific pcr qmsp na  (Thermo Fisher)

Journal: American Journal of Translational Research

Article Title: Upregulation of SPI1 during myocardial infarction aggravates cardiac tissue injury and disease progression through activation of the TLR4/NFκB axis

doi:

Figure Lengend Snippet: High expression of SPI1 in MI was attributed to low level of DNA methylation. (A) Location of SPI1 promoter in mouse genome; (B) CpG island on the SPI1 promoter; (C, D) DNA methylation level of SPI1 in mouse cardiac tissues (C) and in HL-1 cells (D) examined by qMSP; (E) Binding peaks between DNMTs with SPI1 promoter according to the ChIP-seq analysis; (F, G) Abundance of DNMTs in the promoter region of SPI1 in mouse cardiac tissues (F) and HL-1 cells (G) examined by ChIP-qPCR; (H) DNA methylation in SPI1 promoter in HL-1 cells after 5-Azadc treatment examined by qMSP; (I, J) mRNA (I) and protein (J) levels of SPI1 in HL-1 cells after 5-Azadc treatment determined by RT-qPCR and western blot analysis. For cellular experiments, repetition = 3. All data are presented as mean ± SD. Differences were analyzed by unpaired t test (C, D, H, and I) or two-way ANOVA (F and G). *P < 0.05 vs. the sham group; #P < 0.05 vs. the control group; &P < 0.05 vs. the DMSO group.

Article Snippet: Quantitative methylation specific PCR (qMSP) DNA from tissue sections was extracted and isolated by an Ion AmpliSeqTM Direct FFPE DNA Kit (Thermo Fisher Scientific).

Techniques: Expressing, DNA Methylation Assay, Binding Assay, ChIP-sequencing, ChIP-qPCR, Quantitative RT-PCR, Western Blot, Control

Journal: American Journal of Translational Research

Article Title: Upregulation of SPI1 during myocardial infarction aggravates cardiac tissue injury and disease progression through activation of the TLR4/NFκB axis

doi:

Figure Lengend Snippet: Diagram presentation for the molecules of action. During MI, the DNA methylation of SPI1 is significantly reduced, which leads to SPI1 transcriptional activation. SPI1 binds to the TLR4 promoter and promotes TLR4 transcription, which augments inflammatory responses and myocardial fibrosis by inducing NFκB phosphorylation.

Article Snippet: Quantitative methylation specific PCR (qMSP) DNA from tissue sections was extracted and isolated by an Ion AmpliSeqTM Direct FFPE DNA Kit (Thermo Fisher Scientific).

Techniques: DNA Methylation Assay, Activation Assay, Phospho-proteomics

Journal: Developmental cell

Article Title: A Yap-Myc-Sox2-p53 Regulatory Network Dictates Metabolic Homeostasis and Differentiation in Kras-driven Pancreatic Ductal Adenocarcinomas

doi: 10.1016/j.devcel.2019.07.022

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Bisulfate Conversion and quantitative Methylation-Specific PCR (qMSP) Genomic DNA was purified from the pancreatic tissues of WT, KF, and KYYF using PrepEase Genomic DNA isolation kit (Affymetrix, Cleveland, Ohio) according to the manufacture’s instruction.

Techniques: Recombinant, Viability Assay, Imaging, TUNEL Assay, Methylation, Plasmid Preparation, Polymer, cDNA Synthesis, SYBR Green Assay, Bicinchoninic Acid Protein Assay, DNA Extraction, Amplification, Knock-Out, CRISPR, Software