Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: KDR and FLT4 in HDMECs, HUVECs and leukemia cell lines . KDR and FLT4 mRNA expression levels were determined by quantitative real-time PCR and indicated underneath the cell line name. TBP expression was used as endogenous control and cell lines CMK (KDR) and HEL (FLT4) were used for normalization. KDR protein expression levels--determined by Western blot analysis--are higher in HDMECs and HUVECs than in positive cell lines (note 1:10 lysate dilution in primary cells). FLT4 protein expression is higher in HDMECs than in HUVECs (note 1:10 lysate dilution in HDMECs) and in positive cell lines. Protein expression corresponds to mRNA expression pattern.

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

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

Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: Promoter methylation status and expression levels of KDR and FLT4

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

Techniques: Methylation, Expressing

Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: Bisulfite sequencing of the KDR promoter . A CpG island is located between -1231 and 1125 relative to the ATG codon of KDR . The 3' part of the KDR promoter region and exon 1 (612 bp, 53 CpG sites) were sequenced after bisulfite conversion of DNA from cell lines DOHH-2 ( KDR negative) and HEL ( KDR positive) as well as from HDMECs and HUVECs. Each line depicts a sequenced clone representing the methylation status of an individual allele. CpGs are represented as open dots (if unmethylated) or filled dots (if methylated). Results of qRT-PCR and methylation specific PCR (MSP) are shown on the left hand side. M: signal in M-PCR; U: signal in U-PCR.

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

Techniques: Methylation Sequencing, Methylation, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: Methylation status of KDR and FLT4 in cell lines and primary cells . The methylation status of KDR and FLT4 in leukemia cell lines and in HDMECs and HUVECs was analyzed by MSP after bisulfite conversion of the DNA. Agarose gels of KDR and FLT4 M- and U-PCR are shown. A complete list of results is shown in Table 2. NTC, non template control.

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

Techniques: Methylation, Control

Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: Effect of 5-Aza-dC on expression of KDR and FLT4

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

Techniques: Expressing

Journal: BMC Cancer

Article Title: DNA methylation regulates expression of VEGF-R2 ( KDR ) and VEGF-R3 ( FLT4 )

doi: 10.1186/1471-2407-12-19

Figure Lengend Snippet: Expression of KDR and FLT4 on HDMECs and HUVECs . Flow cytometry analysis for KDR, FLT4, the lymphatic vessel marker podoplanin and the panendothelial marker CD31. Note that HDMECs and HUVECs show comparable KDR expression levels, while FLT4 is stronger in lymphatic vessel cells (HDMECs) than in blood vessel cells (HUCECs). The non-filled peak shows the corresponding isotype control.

Article Snippet: To confirm the methylation status of the FLT4 and KDR promoters, genomic DNA was bisulfite converted according to the manufacturer's instructions (Qiagen).

Techniques: Expressing, Flow Cytometry, Marker, Control