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Bisulfite PCR sequencing (4 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the NTN1 CpG island (CGi) was inversely correlated with its expression. Pearson correlation, P = 0.008, r = −0.55, n = 18. Bisulfite PCR sequencing (3 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the DAPK1 CGi was inversely correlated with its expression. Pearson correlation, P = 0.003, r = −0.66, n = 22. Tissue <t>microarrays</t> (70 paraffin embedded sections) from human breast carcinomas were immuno‐stained with antibodies against DAPK1, UNC5B, and netrin‐1. Samples were classified in quartiles according to the level of netrin‐1 expression. The levels of DAPK1 and UNC5B expression (index constructed from the percentage of sections exhibiting a positive staining) in the first and fourth quartiles of netrin‐1 expressing groups were compared using a chi‐squared test. Representative staining corresponding to low and high levels of expression is shown for each antibody, and expression levels were determined from the percentage of sections exhibiting a positive staining. As a control, staining of the samples using a non‐related isotype antibody was performed. Quantification of the presence or absence of alterations in gene A was associated with the presence or absence of alterations in gene B in human breast tumors which was determined from cBioPortal web site, z ‐score threshold ± 2.2 Individual methylation plots. Red rectangles represent methylated CpG and blue rectangles unmethylated CpG. Mean DNA methylation inhibition of DAPK1 and NTN1 in MDA‐MB‐231 and HMLER cells upon DAC treatment.
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Bisulfite PCR sequencing (4 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the NTN1 CpG island (CGi) was inversely correlated with its expression. Pearson correlation, P = 0.008, r = −0.55, n = 18. Bisulfite PCR sequencing (3 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the DAPK1 CGi was inversely correlated with its expression. Pearson correlation, P = 0.003, r = −0.66, n = 22. Tissue microarrays (70 paraffin embedded sections) from human breast carcinomas were immuno‐stained with antibodies against DAPK1, UNC5B, and netrin‐1. Samples were classified in quartiles according to the level of netrin‐1 expression. The levels of DAPK1 and UNC5B expression (index constructed from the percentage of sections exhibiting a positive staining) in the first and fourth quartiles of netrin‐1 expressing groups were compared using a chi‐squared test. Representative staining corresponding to low and high levels of expression is shown for each antibody, and expression levels were determined from the percentage of sections exhibiting a positive staining. As a control, staining of the samples using a non‐related isotype antibody was performed. Quantification of the presence or absence of alterations in gene A was associated with the presence or absence of alterations in gene B in human breast tumors which was determined from cBioPortal web site, z ‐score threshold ± 2.2 Individual methylation plots. Red rectangles represent methylated CpG and blue rectangles unmethylated CpG. Mean DNA methylation inhibition of DAPK1 and NTN1 in MDA‐MB‐231 and HMLER cells upon DAC treatment.

Journal: EMBO Molecular Medicine

Article Title: Inhibition of DNA methylation promotes breast tumor sensitivity to netrin‐1 interference

doi: 10.15252/emmm.201505945

Figure Lengend Snippet: Bisulfite PCR sequencing (4 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the NTN1 CpG island (CGi) was inversely correlated with its expression. Pearson correlation, P = 0.008, r = −0.55, n = 18. Bisulfite PCR sequencing (3 CpGs analyzed, region: light gray boxes in Fig A) indicated that DNA methylation of the DAPK1 CGi was inversely correlated with its expression. Pearson correlation, P = 0.003, r = −0.66, n = 22. Tissue microarrays (70 paraffin embedded sections) from human breast carcinomas were immuno‐stained with antibodies against DAPK1, UNC5B, and netrin‐1. Samples were classified in quartiles according to the level of netrin‐1 expression. The levels of DAPK1 and UNC5B expression (index constructed from the percentage of sections exhibiting a positive staining) in the first and fourth quartiles of netrin‐1 expressing groups were compared using a chi‐squared test. Representative staining corresponding to low and high levels of expression is shown for each antibody, and expression levels were determined from the percentage of sections exhibiting a positive staining. As a control, staining of the samples using a non‐related isotype antibody was performed. Quantification of the presence or absence of alterations in gene A was associated with the presence or absence of alterations in gene B in human breast tumors which was determined from cBioPortal web site, z ‐score threshold ± 2.2 Individual methylation plots. Red rectangles represent methylated CpG and blue rectangles unmethylated CpG. Mean DNA methylation inhibition of DAPK1 and NTN1 in MDA‐MB‐231 and HMLER cells upon DAC treatment.

Article Snippet: Tissue microarrays of paraffin embedded breast tumor sections were obtained from Super Bio Chips (Cliniscience, Nanterre, France).

Techniques: Sequencing, DNA Methylation Assay, Expressing, Staining, Construct, Methylation, Inhibition