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Hcy-induced elevation of EZH2 is responsible for CFTR promoter H3K27me3. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy and adenovirus harboring EZH2 small interfering RNA (si-EZH2) respectively for 24 h. a The H3K27me1, 2, 3 levels on the CFTR promoter were examined in liver of mice by <t>ChIP-PCR.</t> b The levels of H3K27me3 on the CFTR promoter were examined by ChIP-PCR in hepatocytes treated with 100μmol/L Hcy and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. c The H3K27me3 levels in the CFTR promoter of hepatocytes treated with different concentration of agonist of H3K27me3 (1, 5, 10 μmol/L EPZ005687). d , e <t>qRT-PCR</t> and western blot were used to detect the mRNA and protein expression of CFTR in the hepatic cells treated with 10 μmol/L EPZ005687 and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. f , g The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in HL-7702 cells exposed to 100μmol/L Hcy for 24 h. ( H - I ) The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in hepatic cells infected with adenovirus si-EZH2. ( J ) The level of H3K27me3 on CFTR promoter were detected by ChIP-PCR in hepatic cells infected with adenovirus si-EZH2. ( K ) The mRNA and protein expression of CFTR were detected by western blot in hepatic cells infected with adenovirus si-EZH2. ( L ) The protein expression of autophagy related protein factors p62, BECN1, LC3 and Atg12 were detected by western blot in hepatocytes infected with adenovirus (Ad-CFTR, si-EZH2, Ad-CFTR and si-EZH2). Mean ± SD of three experiments performed in triplicate. * P
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1) Product Images from "Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver"

Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver

Journal: Cell Death & Disease

doi: 10.1038/s41419-017-0216-z

Hcy-induced elevation of EZH2 is responsible for CFTR promoter H3K27me3. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy and adenovirus harboring EZH2 small interfering RNA (si-EZH2) respectively for 24 h. a The H3K27me1, 2, 3 levels on the CFTR promoter were examined in liver of mice by ChIP-PCR. b The levels of H3K27me3 on the CFTR promoter were examined by ChIP-PCR in hepatocytes treated with 100μmol/L Hcy and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. c The H3K27me3 levels in the CFTR promoter of hepatocytes treated with different concentration of agonist of H3K27me3 (1, 5, 10 μmol/L EPZ005687). d , e qRT-PCR and western blot were used to detect the mRNA and protein expression of CFTR in the hepatic cells treated with 10 μmol/L EPZ005687 and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. f , g The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in HL-7702 cells exposed to 100μmol/L Hcy for 24 h. ( H - I ) The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in hepatic cells infected with adenovirus si-EZH2. ( J ) The level of H3K27me3 on CFTR promoter were detected by ChIP-PCR in hepatic cells infected with adenovirus si-EZH2. ( K ) The mRNA and protein expression of CFTR were detected by western blot in hepatic cells infected with adenovirus si-EZH2. ( L ) The protein expression of autophagy related protein factors p62, BECN1, LC3 and Atg12 were detected by western blot in hepatocytes infected with adenovirus (Ad-CFTR, si-EZH2, Ad-CFTR and si-EZH2). Mean ± SD of three experiments performed in triplicate. * P
Figure Legend Snippet: Hcy-induced elevation of EZH2 is responsible for CFTR promoter H3K27me3. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy and adenovirus harboring EZH2 small interfering RNA (si-EZH2) respectively for 24 h. a The H3K27me1, 2, 3 levels on the CFTR promoter were examined in liver of mice by ChIP-PCR. b The levels of H3K27me3 on the CFTR promoter were examined by ChIP-PCR in hepatocytes treated with 100μmol/L Hcy and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. c The H3K27me3 levels in the CFTR promoter of hepatocytes treated with different concentration of agonist of H3K27me3 (1, 5, 10 μmol/L EPZ005687). d , e qRT-PCR and western blot were used to detect the mRNA and protein expression of CFTR in the hepatic cells treated with 10 μmol/L EPZ005687 and 100 μmol/L Hcy plus 10 μmol/L EPZ005687. f , g The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in HL-7702 cells exposed to 100μmol/L Hcy for 24 h. ( H - I ) The mRNA and protein expression of EZH2 were detected by qRT-PCR and western blot in hepatic cells infected with adenovirus si-EZH2. ( J ) The level of H3K27me3 on CFTR promoter were detected by ChIP-PCR in hepatic cells infected with adenovirus si-EZH2. ( K ) The mRNA and protein expression of CFTR were detected by western blot in hepatic cells infected with adenovirus si-EZH2. ( L ) The protein expression of autophagy related protein factors p62, BECN1, LC3 and Atg12 were detected by western blot in hepatocytes infected with adenovirus (Ad-CFTR, si-EZH2, Ad-CFTR and si-EZH2). Mean ± SD of three experiments performed in triplicate. * P

Techniques Used: Knock-Out, Mouse Assay, Small Interfering RNA, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Concentration Assay, Quantitative RT-PCR, Western Blot, Expressing, Infection

Hcy induces hepatic autophagy in CBS +/- mice. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy (100 μmol/L) for 24 h. a The concentration of Hcy in plasma of mice was measured by automatic biochemical analyzer. b Contents of ALT and AST in serum of CBS +/- mice were analyzed using automatic biochemical analyzer. c Hematoxylin and eosin (H E) and Oil Red O staining of CBS +/- mice liver. d Transmission electron microscope (TEM) was used to analyse cell autophagy in liver tissues of CBS +/− mice. The arrows indicate the double-membrane vacuoles digesting organelles or cytosolic contents. e and f mRNA and protein expression of p62, BECN1, LC3 and Atg12 in the liver tissue of CBS +/− mice by qRT-PCR and western blot, respectively. g Hepatocytes were treated with different concentrations of Hcy (50–500 μmol/L) for 24 h before MTT assay. h The activities of AST and ALT in hepatocytes after treatment with Hcy were detected by ELISA. ( I ) Confocal fluorescent microscopy analysis of hepatocytes overexpressing mRFP-GFP-LC3, treated with 100 µmol/L Hcy for 24 h. Quantification of mean red and green fluorescent puncta of at least 10 cells per condition is shown. The efficiency of transfection was also shown. ( J ) and ( K ) mRNA and protein expression of p62, BECN1, LC3 and Atg12 in hepatic cells treated with 100 µmol/L L-Hcy by qRT-PCR and western blot. Densitometry analysis of the proteins was performed for each sample (mean ± s.d.). * P
Figure Legend Snippet: Hcy induces hepatic autophagy in CBS +/- mice. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy (100 μmol/L) for 24 h. a The concentration of Hcy in plasma of mice was measured by automatic biochemical analyzer. b Contents of ALT and AST in serum of CBS +/- mice were analyzed using automatic biochemical analyzer. c Hematoxylin and eosin (H E) and Oil Red O staining of CBS +/- mice liver. d Transmission electron microscope (TEM) was used to analyse cell autophagy in liver tissues of CBS +/− mice. The arrows indicate the double-membrane vacuoles digesting organelles or cytosolic contents. e and f mRNA and protein expression of p62, BECN1, LC3 and Atg12 in the liver tissue of CBS +/− mice by qRT-PCR and western blot, respectively. g Hepatocytes were treated with different concentrations of Hcy (50–500 μmol/L) for 24 h before MTT assay. h The activities of AST and ALT in hepatocytes after treatment with Hcy were detected by ELISA. ( I ) Confocal fluorescent microscopy analysis of hepatocytes overexpressing mRFP-GFP-LC3, treated with 100 µmol/L Hcy for 24 h. Quantification of mean red and green fluorescent puncta of at least 10 cells per condition is shown. The efficiency of transfection was also shown. ( J ) and ( K ) mRNA and protein expression of p62, BECN1, LC3 and Atg12 in hepatic cells treated with 100 µmol/L L-Hcy by qRT-PCR and western blot. Densitometry analysis of the proteins was performed for each sample (mean ± s.d.). * P

Techniques Used: Mouse Assay, Knock-Out, Concentration Assay, AST Assay, Staining, Transmission Assay, Microscopy, Transmission Electron Microscopy, Expressing, Quantitative RT-PCR, Western Blot, MTT Assay, Enzyme-linked Immunosorbent Assay, Transfection

Hcy promotes the autophagy of hepatocytes by downregulation of cystic fibrosis trans membrane conductance regulator ( CFTR ). Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy, VX-770 and CFTR(inh)-172, respectively, for 24 h. a , b qRT-PCR and western blot detected the mRNA and protein expression of CFTR in the liver of CBS +/- mice. c , d qRT-PCR and western blot detected the mRNA and protein expression of CFTR in HL-7702 cells treated with 100 μmol/L Hcy. e , f Effect of CFTR activation on the expression of autophagy related proteins p62, BECN1, LC3-II/I and Atg12 in hepatocytes treated with VX-770. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. g , h Effect of CFTR inhibition on the expression of autophagy related proteins p62, BECN1, LC3-II/I and Atg12 in hepatocytes treated with CFTR(inh)-172. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. i , j, k An adenoviral vector carrying either a scrambled sequence or CFTR plasmid infected hepatocytes and protein expression of CFTR was detected by western blot, and the effect of CFTR overexpression and activation on the ratio of LC3-II/I and the expression of autophagy related proteins p62, BECN1 and Atg12 was examined in hepatocytes infected with Ad-CFTR. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. Results represent the means ± s.d. of independent experiments. * P
Figure Legend Snippet: Hcy promotes the autophagy of hepatocytes by downregulation of cystic fibrosis trans membrane conductance regulator ( CFTR ). Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy, VX-770 and CFTR(inh)-172, respectively, for 24 h. a , b qRT-PCR and western blot detected the mRNA and protein expression of CFTR in the liver of CBS +/- mice. c , d qRT-PCR and western blot detected the mRNA and protein expression of CFTR in HL-7702 cells treated with 100 μmol/L Hcy. e , f Effect of CFTR activation on the expression of autophagy related proteins p62, BECN1, LC3-II/I and Atg12 in hepatocytes treated with VX-770. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. g , h Effect of CFTR inhibition on the expression of autophagy related proteins p62, BECN1, LC3-II/I and Atg12 in hepatocytes treated with CFTR(inh)-172. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. i , j, k An adenoviral vector carrying either a scrambled sequence or CFTR plasmid infected hepatocytes and protein expression of CFTR was detected by western blot, and the effect of CFTR overexpression and activation on the ratio of LC3-II/I and the expression of autophagy related proteins p62, BECN1 and Atg12 was examined in hepatocytes infected with Ad-CFTR. The mRNA and protein changes were detected by qRT-PCR and western blot respectively. Results represent the means ± s.d. of independent experiments. * P

Techniques Used: Knock-Out, Mouse Assay, Quantitative RT-PCR, Western Blot, Expressing, Activation Assay, Inhibition, Plasmid Preparation, Sequencing, Infection, Over Expression

Hcy induces liver autophagy by downregulation of CFTR expression via DNA methylation. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy and adenovirus harboring DNMT1 small interfering RNA (si-DNMT1) respectively for 24 h. a A schematic drawing of the putative CpG islands in the 5’-flank region of CFTR . (B, C) The total methylation rate of CpG rich region (−572 bp/−262 bp) within the proximal promoter of the CFTR gene was detected by bisulfite-sequencing PCR (BSP) method in liver of CBS +/- mice and ( d , E ) HL-7702 cells. Black and white circles represent methylated and unmethylated CpGs respectively. f qRT-PCR and western blot were used to detect the mRNA and protein expression of CFTR in HL-7702 cells treated with Hcy (100 μmol/L) or Hcy (100 μmol/L) plus 5-azacytidine (AZC) (10 μmol/L). g , h The mRNA and protein expression of DNMTs in liver tissues of mice were detected by qRT-PCR and western blot. i The mRNA and protein expression of DNMT1 were detected by qRT-PCR and western blot in hepatic cells infected with adenovirus si-DNMT1. j Bisulfite sequencing detected the DNA methylation of CFTR in HL-7702 cells infected with the adenovirus si-DNMT1. k Western blot was performed to detect the protein expression of autophagy related proteins p62, BECN1, LC3 and Atg12 in hepatocytes infected with adenovirus CFTR and adenovirus si-DNMT1. Mean ± s.d. of three experiments performed in triplicate. * P
Figure Legend Snippet: Hcy induces liver autophagy by downregulation of CFTR expression via DNA methylation. Eight to 10 weeks old cystathionine b-synthase ( CBS ) heterozygous knockout mice ( CBS +/− ) were fed with regular mice chow and water ad libitum. Human hepatocytes HL-7702 were treated with L-Hcy and adenovirus harboring DNMT1 small interfering RNA (si-DNMT1) respectively for 24 h. a A schematic drawing of the putative CpG islands in the 5’-flank region of CFTR . (B, C) The total methylation rate of CpG rich region (−572 bp/−262 bp) within the proximal promoter of the CFTR gene was detected by bisulfite-sequencing PCR (BSP) method in liver of CBS +/- mice and ( d , E ) HL-7702 cells. Black and white circles represent methylated and unmethylated CpGs respectively. f qRT-PCR and western blot were used to detect the mRNA and protein expression of CFTR in HL-7702 cells treated with Hcy (100 μmol/L) or Hcy (100 μmol/L) plus 5-azacytidine (AZC) (10 μmol/L). g , h The mRNA and protein expression of DNMTs in liver tissues of mice were detected by qRT-PCR and western blot. i The mRNA and protein expression of DNMT1 were detected by qRT-PCR and western blot in hepatic cells infected with adenovirus si-DNMT1. j Bisulfite sequencing detected the DNA methylation of CFTR in HL-7702 cells infected with the adenovirus si-DNMT1. k Western blot was performed to detect the protein expression of autophagy related proteins p62, BECN1, LC3 and Atg12 in hepatocytes infected with adenovirus CFTR and adenovirus si-DNMT1. Mean ± s.d. of three experiments performed in triplicate. * P

Techniques Used: Expressing, DNA Methylation Assay, Knock-Out, Mouse Assay, Small Interfering RNA, Methylation, Methylation Sequencing, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Infection

CFTR alters the levels of liver injury biomarker and autophagy in CBS +/ − mice. CBS +/− mice injected with lentivirus Lv-CFTR or Lv-GFP through the tail vein (see Materials and Methods). a CFTR mRNA and protein levels were determined by qRT-PCR and western blot respectively. Data are presented as mean ± s.d. relative to Lv-GFP-injected mice. b Circulating levels of ALT and AST were analyzed. Results are means ± s.d. ( n = 6). * P
Figure Legend Snippet: CFTR alters the levels of liver injury biomarker and autophagy in CBS +/ − mice. CBS +/− mice injected with lentivirus Lv-CFTR or Lv-GFP through the tail vein (see Materials and Methods). a CFTR mRNA and protein levels were determined by qRT-PCR and western blot respectively. Data are presented as mean ± s.d. relative to Lv-GFP-injected mice. b Circulating levels of ALT and AST were analyzed. Results are means ± s.d. ( n = 6). * P

Techniques Used: Biomarker Assay, Mouse Assay, Injection, Quantitative RT-PCR, Western Blot, AST Assay

2) Product Images from "Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer"

Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer

Journal: Cell Death & Disease

doi: 10.1038/s41419-018-0908-z

Knockdown of NEAT1 inhibits ovarian tumor growth in vivo. a NEAT1-knockdown stable cell lines were constructed, and the knockdown efficiency was tested by qRT-PCR. b Downregulation of NEAT1 expression attenuated tumor growth in nude mice. c , d The effect of NEAT1 on EOC tumor growth was evaluated based on the tumor volumes and tumor weights in the two groups. e An immunofluorescence assay was used to detect the Ki-67 expression levels in frozen sections. Scale bar is 50 μm. * P
Figure Legend Snippet: Knockdown of NEAT1 inhibits ovarian tumor growth in vivo. a NEAT1-knockdown stable cell lines were constructed, and the knockdown efficiency was tested by qRT-PCR. b Downregulation of NEAT1 expression attenuated tumor growth in nude mice. c , d The effect of NEAT1 on EOC tumor growth was evaluated based on the tumor volumes and tumor weights in the two groups. e An immunofluorescence assay was used to detect the Ki-67 expression levels in frozen sections. Scale bar is 50 μm. * P

Techniques Used: In Vivo, Stable Transfection, Construct, Quantitative RT-PCR, Expressing, Mouse Assay, Immunofluorescence

NEAT1 is upregulated in HGSOC and is correlated with patient prognosis. a NEAT1 expression levels in HGSOC and unpaired normal ovarian tissues determined using the NEAT1 primer (which can detect both transcripts). b NEAT1 expression in HGSOC and normal tissues was determined using the NEAT1-2 primer (which can detect the long transcript). β-actin was used as an endogenous control to normalize the data. c A total of 75 HGSOC patients were divided into high- and low-expression groups based on the median expression value. d Kaplan–Meier analysis of overall survival in all patients with HGSOC according to NEAT1 expression. e Kaplan–Meier curves for PFS of patients based on NEAT1 expression. f NEAT1 expression was quantitated in seven EOC cell lines using qRT-PCR
Figure Legend Snippet: NEAT1 is upregulated in HGSOC and is correlated with patient prognosis. a NEAT1 expression levels in HGSOC and unpaired normal ovarian tissues determined using the NEAT1 primer (which can detect both transcripts). b NEAT1 expression in HGSOC and normal tissues was determined using the NEAT1-2 primer (which can detect the long transcript). β-actin was used as an endogenous control to normalize the data. c A total of 75 HGSOC patients were divided into high- and low-expression groups based on the median expression value. d Kaplan–Meier analysis of overall survival in all patients with HGSOC according to NEAT1 expression. e Kaplan–Meier curves for PFS of patients based on NEAT1 expression. f NEAT1 expression was quantitated in seven EOC cell lines using qRT-PCR

Techniques Used: Expressing, Quantitative RT-PCR

Attenuation of NEAT1 expression inhibits cell proliferation, invasion, and migration in vitro. a The EOC cell lines OVCAR3 and A2780 were transfected with siRNAs, and the efficiency of knockdown was verified by qRT-PCR using two NEAT1 primers. b CCK-8 assays were performed to determine the proliferation of NEAT1-knockdown cells. c The cell colony formation ability of OVCAR3 and A2780 cells was assessed to determine the effects of NEAT1 knockdown on cell growth. d The cell invasion potential of the OVCAR3 and A2780 cells was assessed using a Transwell assay. Scale bar is 50 μm. e The cell migration ability was evaluated using a wound-healing assay; images of OVCAR3 and A2780 cells were taken at 0 and 36 h postscratch. Scale bar is 200 μm. f Western blotting analysis was used to determine the MMP2, MMP9, N-cadherin, and E-cadherin expression levels. Actin was used as a reference. The data are shown as the mean ± SD of three replicates; * P
Figure Legend Snippet: Attenuation of NEAT1 expression inhibits cell proliferation, invasion, and migration in vitro. a The EOC cell lines OVCAR3 and A2780 were transfected with siRNAs, and the efficiency of knockdown was verified by qRT-PCR using two NEAT1 primers. b CCK-8 assays were performed to determine the proliferation of NEAT1-knockdown cells. c The cell colony formation ability of OVCAR3 and A2780 cells was assessed to determine the effects of NEAT1 knockdown on cell growth. d The cell invasion potential of the OVCAR3 and A2780 cells was assessed using a Transwell assay. Scale bar is 50 μm. e The cell migration ability was evaluated using a wound-healing assay; images of OVCAR3 and A2780 cells were taken at 0 and 36 h postscratch. Scale bar is 200 μm. f Western blotting analysis was used to determine the MMP2, MMP9, N-cadherin, and E-cadherin expression levels. Actin was used as a reference. The data are shown as the mean ± SD of three replicates; * P

Techniques Used: Expressing, Migration, In Vitro, Transfection, Quantitative RT-PCR, CCK-8 Assay, Transwell Assay, Wound Healing Assay, Western Blot

LIN28B enhances the stability of NEAT1. a Immunoblotting to evaluate the LIN28B protein levels after NEAT1 downregulation in OVCAR3 and A2780 cells. b The qRT-PCR analysis of LIN28B interference in EOC cells. c The EOC cell lines A2780 and OVCAR3 were transfected with either LIN28B or a vector control, and LIN28B overexpression was verified by qRT-PCR. Relative RNA levels of NEAT1 in OVCAR3 and A2780 cells with LIN28B knockdown ( d ) or overexpression ( e ) based on qPCR. f The half-life of NEAT1 after treatment with 2.5 μM actinomycin D for the indicated times with LIN28B overexpression in A2780 cells. * P
Figure Legend Snippet: LIN28B enhances the stability of NEAT1. a Immunoblotting to evaluate the LIN28B protein levels after NEAT1 downregulation in OVCAR3 and A2780 cells. b The qRT-PCR analysis of LIN28B interference in EOC cells. c The EOC cell lines A2780 and OVCAR3 were transfected with either LIN28B or a vector control, and LIN28B overexpression was verified by qRT-PCR. Relative RNA levels of NEAT1 in OVCAR3 and A2780 cells with LIN28B knockdown ( d ) or overexpression ( e ) based on qPCR. f The half-life of NEAT1 after treatment with 2.5 μM actinomycin D for the indicated times with LIN28B overexpression in A2780 cells. * P

Techniques Used: Quantitative RT-PCR, Transfection, Plasmid Preparation, Over Expression, Real-time Polymerase Chain Reaction

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Amplification:

Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver
Article Snippet: Quantitative real-time PCR (qRT-PCR) for CFTR , DNMT1 and EZH2 genes was performed using an FTC3000 qRT-PCR detection system as described previously , . qRT-PCR kits were from Takara Biotechnology Co., Ltd. (Dalian, China). .. The first-strand cDNA reaction (0.5 μL) was subjected to qRT-PCR amplification using gene-specific primers (Table ).

Synthesized:

Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer
Article Snippet: Reverse transcription (RT) and qRT-PCR kits (Takara, Dalian, China) were utilized to evaluate the mRNA expression levels of the indicated genes. .. PCR primers were designed and synthesized using a primer design tool ( http://www.ncbi.nlm.nih.gov/tools/primer-blast/ ); the primer sequences are listed in Supplementary Table .

Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver
Article Snippet: Quantitative real-time PCR (qRT-PCR) for CFTR , DNMT1 and EZH2 genes was performed using an FTC3000 qRT-PCR detection system as described previously , . qRT-PCR kits were from Takara Biotechnology Co., Ltd. (Dalian, China). .. All primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). qRT-PCR reaction was performed with an initial denaturation at 95 °C for 10 min followed by 40 cycles of denaturation at 95 °C for 20 s, annealing at 58 °C for 30 s, and extension at 72 °C for 30 s, and, for a final step, a melting curve of 94 °C for 90 s, 60 °C for 180 s, and 94 °C for 10 s. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as the invariant control; mRNA expression was determined by 2−ΔΔCT method.

Isolation:

Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer
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CCK-8 Assay:

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Quantitative RT-PCR:

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Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer
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Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver
Article Snippet: .. Quantitative real-time PCR (qRT-PCR) for CFTR , DNMT1 and EZH2 genes was performed using an FTC3000 qRT-PCR detection system as described previously , . qRT-PCR kits were from Takara Biotechnology Co., Ltd. (Dalian, China). .. The first-strand cDNA reaction (0.5 μL) was subjected to qRT-PCR amplification using gene-specific primers (Table ).

Real-time Polymerase Chain Reaction:

Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer
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Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver
Article Snippet: .. Quantitative real-time PCR (qRT-PCR) for CFTR , DNMT1 and EZH2 genes was performed using an FTC3000 qRT-PCR detection system as described previously , . qRT-PCR kits were from Takara Biotechnology Co., Ltd. (Dalian, China). .. The first-strand cDNA reaction (0.5 μL) was subjected to qRT-PCR amplification using gene-specific primers (Table ).

Expressing:

Article Title: Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer
Article Snippet: .. Reverse transcription (RT) and qRT-PCR kits (Takara, Dalian, China) were utilized to evaluate the mRNA expression levels of the indicated genes. .. PCR primers were designed and synthesized using a primer design tool ( http://www.ncbi.nlm.nih.gov/tools/primer-blast/ ); the primer sequences are listed in Supplementary Table .

Article Title: Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver
Article Snippet: Quantitative real-time PCR (qRT-PCR) for CFTR , DNMT1 and EZH2 genes was performed using an FTC3000 qRT-PCR detection system as described previously , . qRT-PCR kits were from Takara Biotechnology Co., Ltd. (Dalian, China). .. All primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). qRT-PCR reaction was performed with an initial denaturation at 95 °C for 10 min followed by 40 cycles of denaturation at 95 °C for 20 s, annealing at 58 °C for 30 s, and extension at 72 °C for 30 s, and, for a final step, a melting curve of 94 °C for 90 s, 60 °C for 180 s, and 94 °C for 10 s. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as the invariant control; mRNA expression was determined by 2−ΔΔCT method.

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Polymerase Chain Reaction:

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  • 90
    TaKaRa nucleospin rna extraction kit
    Nucleospin Rna Extraction Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nucleospin rna extraction kit - by Bioz Stars, 2020-01
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    TaKaRa lenti x qrt pcr titration kit
    Lenti X Qrt Pcr Titration Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lenti x qrt pcr titration kit - by Bioz Stars, 2020-01
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    89
    TaKaRa retro x qrt pcr titration kit
    Retro X Qrt Pcr Titration Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 89 stars, based on 10 article reviews
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    retro x qrt pcr titration kit - by Bioz Stars, 2020-01
    89/100 stars
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