Journal: Molecular Syndromology

Article Title: Differences in Copy Number Variation between Discordant Monozygotic Twins as a Model for Exploring Chromosomal Mosaicism in Congenital Heart Defects

doi: 10.1159/000335284

Figure Lengend Snippet: Quantitative PCR results of the 3 confirmed CNV differences between the members of twin pair 6. Quantitative PCR results are shown for the affected and unaffected (UA) member of twin pair 6, as well as for 3 unrelated normal controls (2 males (M), 1 female

Article Snippet: To confirm the presence of CNV differences, quantitative polymerase chain reaction (qPCR) was performed on DNA from both twin members and 3 unrelated controls (2 males and 1 female) using the LightCycler® 480 Real-Time PCR system (Roche Applied Science, Belgium, ), following the manufacturer's instructions.

Techniques: Real-time Polymerase Chain Reaction

Journal: FEMS Microbiology Ecology

Article Title: Cold adaptation and replicable microbial community development during long-term low-temperature anaerobic digestion treatment of synthetic sewage

doi: 10.1093/femsec/fiy095

Figure Lengend Snippet: ( A ) qPCR data of Bacterial and Archaeal 16S copy numbers (per g biomass) on the right y-axis from biomass samples (x-axis) throughout the trial corresponding to the ratio of bacteria to archaea (expressed as a percentage) on the left y-axis. ( B ) qPCR data of Bacterial and Archaeal 16S rRNA transcripts copy numbers (per g biomass) on the right y-axis from biomass samples (x-axis) throughout the trial corresponding to the ratio of bacteria to archaea (expressed as a percentage) on the left y-axis.

Article Snippet: Quantitative-polymerase chain reaction Quantitative polymerase chain reaction (qPCR) was carried out for Archaeal and Bacterial domains using DNA and cDNA generated from granular biomass sampled from R1 and R2 and the fixed-film filter as described above. qPCR was performed using a LightCycler 480 (Roche, Manheim, Germany).

Techniques: Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Potential application of injectable chitosan hydrogel treated with siRNA in chronic rhinosinusitis therapy

doi: 10.3892/mmr.2015.4237

Figure Lengend Snippet: (A) Visualization of the uptake of siRNA using fluorescence microscopy. (B) mRNA expression of VEGF, determined using quantitative polymerase chain reaction analysis. (C) Associated VEGF protein level, determined by enzyme-linked immunosorbent assays. * P

Article Snippet: The mRNA expression of target VEGF was analyzed using quantitative polymerase chain reaction (qPCR) with an SYBR Premix ExTM Taq II kit (Takara Bio, Inc.).

Techniques: Fluorescence, Microscopy, Expressing, Real-time Polymerase Chain Reaction

Journal: Biology Open

Article Title: Generation and analysis of knock-in mice carrying pseudohypoaldosteronism type II-causing mutations in the cullin 3 gene

doi: 10.1242/bio.013276

Figure Lengend Snippet: Expression levels of cullin 3 protein and mRNA and proteins of the WNK–OSR1/SPAK–NCC phosphorylation signaling cascade in the kidneys of Cullin 3 G(−1)A/+ knock-in mice. (A) Immunoblots of proteins of the WNK–OSR1/SPAK–NCC signaling cascade in the kidneys of wild-type (WT) and cullin 3 ( Cul3 ) G(−1)A/+ heterozygous knock-in mice. (B) Densitometry analysis. Values are expressed as a ratio of the average signal in WT mice. Cul3 expression levels in Cul3 G(−1)A/+ heterozygous knock-in mice were approximately half that of WT mice. There were no significant differences in proteins expression levels of the WNK–OSR1/SPAK–NCC phosphorylation signaling cascade between Cul3 G(−1)A/+ heterozygous knock-in and WT mice. (C) Quantitative polymerase chain reaction (PCR) analysis of Cul3 mRNA levels. SYBR Green quantitative PCR was used to quantify mRNA levels in the kidneys of WT mice ( n =7) and Cul3 G(−1)A/+ mice ( n =7). WT: wild type. G(−1)A: Cul3 G(−1)A/+ . * P

Article Snippet: Quantitative polymerase chain reaction Quantitative PCR was performed using SYBR Premix Ex Taq™ II (Takara Bio Inc.) and primers specific for Cul3 , using the forward primer F4 and the reverse primer R5 (5′-AGCCCTGGATATAGTCAACAGG-3′).

Techniques: Expressing, Knock-In, Mouse Assay, Western Blot, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, SYBR Green Assay

Journal: Biology Open

Article Title: Generation and analysis of knock-in mice carrying pseudohypoaldosteronism type II-causing mutations in the cullin 3 gene

doi: 10.1242/bio.013276

Figure Lengend Snippet: Expression levels of cullin 3 protein and mRNA and proteins of the WNK-OSR1/SPAK-NCC phosphorylation signaling cascade in the kidneys of cullin 3 T(−6)G/T(−6)G knock-in mice. (A) Immunoblots of proteins of the WNK–OSR1/SPAK–NCC signaling cascade in the kidneys of wild-type (WT) and cullin 3 ( Cul3 ) T( − 6)G/T( − 6)G homozygous knock-in mice. (B) Densitometry analysis. Values are expressed as the ratio of the average signal in WT mice. The level of Cul3 protein expression in Cul3 T( − 6)G/T( − 6)G mice was three-quarters that of WT mice. There were no significant differences in proteins expression levels of the WNK–OSR1/SPAK–NCC phosphorylation signaling cascade between Cul3 T( − 6)G/T( − 6)G and WT mice. (C) Quantitative polymerase chain reaction (PCR) analysis of Cul3 mRNA levels. SYBR Green quantitative PCR was used to quantify mRNA levels in the kidneys of WT mice ( n =7) and Cul3 T( − 6)G/T( − 6)G mice ( n =9). WT: wild-type mice. T(−6)G: Cul3 T( − 6)G/T( − 6)G mice. * P

Article Snippet: Quantitative polymerase chain reaction Quantitative PCR was performed using SYBR Premix Ex Taq™ II (Takara Bio Inc.) and primers specific for Cul3 , using the forward primer F4 and the reverse primer R5 (5′-AGCCCTGGATATAGTCAACAGG-3′).

Techniques: Expressing, Knock-In, Mouse Assay, Western Blot, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, SYBR Green Assay

Journal: Nucleic Acids Research

Article Title: Linker histone partial phosphorylation: effects on secondary structure and chromatin condensation

doi: 10.1093/nar/gkv304

Figure Lengend Snippet: Fold decrements of template DNA of the β-globin and rhodopsin genes after MNase digestion of chromatin samples quantified by qPCR. Dark gray, phosphorylated samples. Light gray, unphosphorylated samples. ( A, C ) Results for the β-globin gene. ( B, D ) Results for the rhodopsin gene. Fold decrements were calculated as 2 ΔCt , using the Ct value of the initial undigested chromatin sample as reference for each gene. Error bars correspond to the interval 2 ΔCt±SD .

Article Snippet: Purified DNA was used as a template for the amplification of β-globin and rhodopsin genes by Quantitative Polymerase Chain Reaction (qPCR), following MIQE guidelines, in a C1000TM thermal cycler (BioRad).

Techniques: Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Effect on the liver cancer cell invasion ability by studying the associations between autophagy and TRAP1 expression

doi: 10.3892/ol.2018.8774

Figure Lengend Snippet: (A) Reverse transcription-quantitative polymerase chain reaction was used to detect the TRAP1 mRNA expression levels in Hep3b2.1–7, HepG2, HepG2.2.15 and Sk-hep1 cells. (B) The protein levels of TRAP1 in the 4 liver cancer cell lines were determined by western blot analysis. β-actin was used as an internal control. (C) WES™ automated capillary-based size sorting system was used to measure the protein level of TRAP1 and β-actin in 4 liver cancer cell lines. (D) The protein levels of TRAP1 were examined in 4 liver cancer cells treated with 0, 5, 10 and 15 µmol/l rapamycin by western blot analysis. β-actin was used as an internal control. *P

Article Snippet: The mRNA and protein expression levels of tumor necrosis factor receptor-associated protein-1 (TRAP-1), Beclin1 and LC3-II/LC3-I were measured by reverse transcription-quantitative polymerase chain reaction, Protein Simple Western and western blot analysis.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Induction of reprogramming of human amniotic epithelial cells into iPS cells by overexpression of Yap, Oct4, and Sox2 through the activation of the Hippo-Yap pathway

doi: 10.3892/etm.2017.4512

Figure Lengend Snippet: Overexpression the three factors, OSY, induced HuAECs to express high levels of ESC markers. (A) The process of induction of HuAEC reprogramming into iPS cells by OSY. (B) Cell morphology of HuAECs; scale, 30 µm. (C) OSY-iPS cells had clone-like morphology; scale, 30 µm. (D) Alkaline phosphatase staining identification of OSY-iPS cells was positive; scale, 30 µm. (E) Semi-quantitative PCR results indicated that OSY-iPS cells expressed high levels of ESC markers (Oct4, Sox2, Nanog, Rex1, and Ssea3/4) and Yap. (F) Immunofluorescence staining results suggested that OSY-iPS cells expressed high levels of Oct4 and SSEA3/4 proteins; scale, 30 µm. (G) Western blot results showed that OSY-iPS cells expressed high levels of Oct4, Sox2, and Yap proteins.

Article Snippet: Quantitative polymerase chain reaction (qPCR) was performed using a RealPlex4 real-time PCR detection system (Eppendorf Co., Ltd., Hamburg, Germany).

Techniques: Over Expression, Staining, Real-time Polymerase Chain Reaction, Immunofluorescence, Western Blot

Journal: Cell Cycle

Article Title: IL-15 and IL-17F are differentially regulated and expressed in mycosis fungoides (MF)

doi: 10.4161/cc.28256

Figure Lengend Snippet: Figure 2. IL-15 expression in malignant T cells. ( A ) Non-malignant (MySi) and malignant (PB2B, MF2000) T cells were cultured for 24 h. RNA was purified from the cells and reversely transcribed to cDNA and analyzed by quantitative PCR to determine

Article Snippet: Total cellular mRNA was purified and reverse transcribed into cDNA (cDNA) as described previously., Quantitative polymerase chain reaction (QPCR) was subsequently performed using the Brilliant II SYBR Green quantitative PCR kit from Stratagene in accordance with the manufacturer’s instructions and the samples analyzed on a Mx3005P (Stratagene).

Techniques: Expressing, Cell Culture, Purification, Real-time Polymerase Chain Reaction