Proposed model.

<p>Panel A illustrates the following: in epithelial cells, <i>CDH1</i>, <i>Mgat3</i> and <i>Mgat5</i> are transcribed. Partial promoter methylation of <i>Mgat3</i> and no methylation of <i>CDH1</i> and <i>Mgat5</i> promoters were observed. The transcription levels of <i>Mgat3</i> generate sufficient GnT-III enzyme levels that catalyze the addition of bisecting GlcNAc structures, specifically on E-cadherin. No information is available concerning the status of the remaining molecules in the adhesion complex (catenins). Panel B illustrates the following: in mesenchymal cells, <i>Mgat3</i>'s promoter is methylated in some <i>CpG</i> sites which were associated with a significant decrease of <i>Mgat3</i> transcription. No significant changes were observed in terms of both promoter methylation status and transcription of <i>CDH1</i> and <i>Mgat5</i>. There was a significant decrease of GnT-III-mediated E-cadherin glycosylation. In reverted epithelial cells, <i>Mgat3</i>'s promoter methylation status returns to its status in original epithelial cells accompanied with a significant increase of <i>Mgat3</i> transcription, in comparison to mesenchymal cells. Concomitantly there was an increased GnT-III-mediated E-cadherin glycosylation, resembling that observed in original epithelial cells.</p

EMT/MET model validation.

<p>Panel A shows the Western blot for E-cadherin. Panel B illustrates the quantification of E-cadherin across the EMT/MET induction (<i>n</i> = 3 biological replicas). Data was normalized for E cells. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033191#s2" target="_blank">Results</a> are described as mean±standard error mean of 3 biological replicas. No significant differences were observed concerning E-cadherin expression (<i>ns</i> stands for non-significant, <i>p</i>>0.05). Panels A and B show that E-cadherin expression is decreased in M cells (in comparison to E cells) and partially recovered in RE cells (in comparison to M cells). Panel C represents the immunofluorescence for E-cadherin during EMT/MET induction (200×). <i>NC</i> stands for negative control (no E-cadherin antibody used). Panel C illustrates the variation of E-cadherin localization during the EMT/MET induction: E cells display the classical E-cadherin expression at the cell membrane; M cells show a decreased expression of E-cadherin which is only observed in some points of intercellular contacts and in the cytoplasm; RE cells display E-cadherin expression in the cell membrane.</p

Pathways of the differentially expressed genes in HL-derived and ALCL-derived cell lines

<p><b>Copyright information:</b></p><p>Taken from "Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines"</p><p>http://www.molecular-cancer.com/content/7/1/2</p><p>Molecular Cancer 2008;7():2-2.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2254646.</p><p></p

Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines-2

<p><b>Copyright information:</b></p><p>Taken from "Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines"</p><p>http://www.molecular-cancer.com/content/7/1/2</p><p>Molecular Cancer 2008;7():2-2.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2254646.</p><p></p

Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines-1

<p><b>Copyright information:</b></p><p>Taken from "Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines"</p><p>http://www.molecular-cancer.com/content/7/1/2</p><p>Molecular Cancer 2008;7():2-2.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2254646.</p><p></p

GnT-III-mediated E-cadherin glycosylation during EMT/MET.

<p>Panel A shows the co-immunofluorescence for E-cadherin and E-PHA (400× for E, M, RE and 630× for M*) illustrating that E-cadherin and bisecting GlcNAc structures co-localize in the cell membrane in E and RE. In mesenchymal cells (M and M*), it was observed a significant decrease in both the expression of E-cadherin and bisecting GlcNAc structures. M cells shows residual E-cadherin expression at the focal points of intercellular contacts (red, E-cadherin) and some green staining (E-PHA reactivity) could be observed in the perinuclear region (Golgi compartment). Immunoprecipitation of E-cadherin followed by E-PHA lectin blot is represented in panel B. Panel C represents the normalization of bisecting GlcNAc structures (E-PHA reactivity) that are modifying E-cadherin. Amounts of N-glycan structures were determined from the ratios of densities of E-PHA reactivity after normalization to E-cadherin. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033191#s2" target="_blank">Results</a> are described as mean ± standard error mean of two biological replicas. Single asterisk corresponds to <i>p</i>≤0.05 and <i>ns</i> stands for non-significant, <i>p</i>>0.05. The modification of E-cadherin with bisecting GlcNAc N-glycan structures in E, M and RE are expressed as the fold increase, compared with the E cells. Panels B and C show that E-cadherin is specifically glycosylated with bisecting GlcNAc structures in E, losing this glycoform in M and recovering again in RE.</p

Expression levels and cellular localization of the product of GnT-III enzyme during EMT/MET induction.

<p>Panel A shows the lectin blot analysis using E-PHA lectin, showing the total expression levels of bisecting GlcNAc structures during EMT/MET. Panel B illustrates the quantification of E-PHA lectin normalized to actin (<i>n</i> = 2 biological replicas). Single asterisk corresponds to <i>p</i>≤0.05 and double asterisks stands for <i>p</i>≤0.001. Bisecting GlcNAc structures significantly decrease when comparing E and M cells and their expression is significantly recovered in RE cells. Panel C represents the immunofluorescence for E-PHA lectin during EMT/MET induction (400×). Bisecting GlcNAc structures are preferentially localized in the cell membrane of E cells. M cells exhibit a clear decrease in expression of the bisecting GlcNAc structures that was only observed in focal areas in the perinuclear region. In RE cells, there was a significant increase in the E-PHA staining showing an increase in the expression levels of bisecting GlcNAc structures that are localized in the cell membrane and in the cytoplasm.</p

<i>Mgat3</i> and Mgat5 RNA expression and methylation status of their predicted promoter-associated CpG islands.

<p>Panel A illustrates the quantification of <i>Mgat3</i> relative mRNA expression (<i>n</i> = 3 biological replicas). Data was normalized for E cells for each biological replica. Single asterisk corresponds to p≤0.05 and double asterisks stands for p≤0.001. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033191#s2" target="_blank">Results</a> are described as mean ± standard error mean of three biological replicas. Panel A shows that <i>Mgat3</i> expression was significantly decreased in M cells (in comparison to E cells) and recovered in RE cells (in comparison to M cells). Schematic representation of <i>Mgat3</i> genomic locus is represented in panel B. White squares correspond to exonic untranslated regions and black squares to exonic translated regions. Black line stands for intronic regions. Grey squares represent the position of the bioinformatically predicted CpG islands (classified as 1 and 2). Panel C and D show the schematic representation of the methylation status of several CpG dinucleotides evaluated within CpG islands 1 (C) and 2 (D) of <i>Mgat3</i> across the EMT/MET experiment (E, M and RE). White circles correspond to unmethylated CpGs, grey circles correspond to partially methylated CpGs, black circles correspond to methylated CpGs, white circles with a question mark correspond to unknown methylation status. Panel C shows methylation pattern alterations across several CpG sites within <i>Mgat3</i>'s CpG island 1 in E, M and RE cells. Panel E illustrates the quantification of Mgat5 relative mRNA expression (<i>n</i> = 3 biological replicas). Same legend as in A applies. No significant variation of Mgat5 RNA expression was observed during EMT/MET. Schematic representation of part of the Mgat5 genomic locus is represented in panel F. Same legend as in panel B applies. Schematic representation of the methylation status of several CpG dinucleotides evaluated within the annotated Mgat5 CpG island is represented in panel G. Same legend as in panels C and D applies. The results showed no variation of the methylation status of Mgat5 promoter during EMT/MET.</p

Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines-3

<p><b>Copyright information:</b></p><p>Taken from "Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines"</p><p>http://www.molecular-cancer.com/content/7/1/2</p><p>Molecular Cancer 2008;7():2-2.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2254646.</p><p></p>reen labeled FISH probes, respectively

Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines-0

<p><b>Copyright information:</b></p><p>Taken from "Sub-megabase resolution tiling (SMRT) array-based comparative genomic hybridization profiling reveals novel gains and losses of chromosomal regions in Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma cell lines"</p><p>http://www.molecular-cancer.com/content/7/1/2</p><p>Molecular Cancer 2008;7():2-2.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2254646.</p><p></p