Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes

Microarray analysis reveals that the specific pattern of gene expression in cardiomyocytes derived from embryonic stem cells reflects the biological, physiological and functional processes occurring in mature cardiomyocytes

Transcriptomic and phenotypic analysis of murine embryonic stem cell derived BMP2+ lineage cells: an insight into mesodermal patterning

Transcriptome analysis of BMP2+ cells in comparison to the undifferentiated BMP2 ES cells and the control population from 7-day old embryoid bodies led to the identification of 479 specifically upregulated and 193 downregulated transcripts

Detection of smooth muscle cells after differentiation of the BMP2cells

Expression of SMA in the BMP2cells detected by qRT-PCR. Microarray relative expression levels of various smooth muscle specific genes in BMP2cells. The expression levels of each gene were normalized with its maximum level set as 100%. Each result is an average of three independent experiments (Additional data file 13). Detection of smooth muscle cells 1 day after plating the BMP2cells (7 + 1 days in total; top left), 8 days after plating with puromycin (7 + 8 days in total; top right), 18 days after plating with puromycin (7 + 18 days in total; bottom left) and 18 days after plating without puromycin (7 + 18 days in total; bottom right).<p><b>Copyright information:</b></p><p>Taken from "Transcriptomic and phenotypic analysis of murine embryonic stem cell derived BMP2lineage cells: an insight into mesodermal patterning"</p><p>http://genomebiology.com/2007/8/9/R184</p><p>Genome Biology 2007;8(9):R184-R184.</p><p>Published online 4 Sep 2007</p><p>PMCID:PMC2375022.</p><p></p

Analysis of osteoblast, chondrocyte and myocyte specific markers in BMP2cells

RT-PCR analysis in BMP2cells. Alizarin stanings on the 18th day after plating the BMP2cells (7 + 18 days in total) and 28 days after plating (7 + 28, 35 days in total). Relative expression levels of the genes presented in (a) as obtained by Affymetrix analysis. The expression levels of each gene were normalized with its maximum level set as 100%. Each result is an average of three independent experiments (Additional data file 13).<p><b>Copyright information:</b></p><p>Taken from "Transcriptomic and phenotypic analysis of murine embryonic stem cell derived BMP2lineage cells: an insight into mesodermal patterning"</p><p>http://genomebiology.com/2007/8/9/R184</p><p>Genome Biology 2007;8(9):R184-R184.</p><p>Published online 4 Sep 2007</p><p>PMCID:PMC2375022.</p><p></p

Hierarchical clustering of probe sets identified as upregulated in α-MHCcells

<p><b>Copyright information:</b></p><p>Taken from "Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes"</p><p>Genome Biology 2007;8(4):R56-R56.</p><p>Published online 11 Apr 2007</p><p>PMCID:PMC1896009.</p><p></p> Shown is a visualization of the hierarchical clustering of probe sets identified as upregulated in the α-myosin heavy chain (MHC)cells with an expression level at least twofold higher than in 15-day-old EBs and in α-MHC embryonic stem cells. Each probe set is represented by a single row of colored boxes; each array is represented by a single column. Rectangles corresponding to intermediately expressed probe sets are colored black, upregulated probe sets are indicated with red of increasing intensity, and weakly expressed probe sets with green of increasing intensity. The dendrogram on the left of the figure represents the similarity matrix of probe sets

Validation of Affymetrix data by quantitative real-time PCR and semi-quantitative PCR analyses

<p><b>Copyright information:</b></p><p>Taken from "Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes"</p><p>Genome Biology 2007;8(4):R56-R56.</p><p>Published online 11 Apr 2007</p><p>PMCID:PMC1896009.</p><p></p> Validation of Affymetrix data by quantitative real-time polymerase chain reaction (PCR) analyses. The fold change was calculated by using the following formula: fold-change = . ΔCof the gene in the sample in which it is expressed lowest is taken as ΔCgene2 to calculate the fold change using the above formula. The resulting fold change is expressed as percentage of the maximum fold change (= 100%) for that particular gene in every assay. Values are expressed as mean ± standard deviation (= 3; technical replicates). Additional validation of Affymetrix data by semi-quantitative reverse transcription (RT)-PCR analyses. Randomly chosen candidate genes to validate Affymetrix data by semi-quantitiative RT-PCR analyses and their relative expression values expressed as percentage of maximum expression for every gene, as obtained from Affymetrix profiling, are given in the table

Enrichment of α-MHCcells isolated from the α-MHCES cell lineage after puromycin treatment

<p><b>Copyright information:</b></p><p>Taken from "Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes"</p><p>Genome Biology 2007;8(4):R56-R56.</p><p>Published online 11 Apr 2007</p><p>PMCID:PMC1896009.</p><p></p> Progressive purification of α-myosin heavy chain (MHC)cardiac cell aggregates after treatment of the 8-day-old embryoid bodies (EBs) with 4 μg/ml puromycin for 7 days. Puromycin containing medium was refreshed every second day. Reverse transcription (RT)-polymerase chain reaction (PCR) analysis of α-MHC expression during EB differentiation (for RT-PCR conditions and primers, see Additional data file 3). Cells from 15-day-old EBs and 15-day-old puromycin purified α-MHCaggregates were dissociated by trypsinization and the purity of the α-MHCcells in the 15-day-old EBs (panel c) and in the 15-day-old α-MHCaggregates (panel d) was examined by fluorescence-activate cell sorting analysis. Characterization of ES cell derived cardiomyocytes by immunocytochemistry. α-MHCcardiomyocytes were dissociated with collagenase B and plated on fibronectin coated coverslips. Enhanced green fluorescent protein (EGFP) expression of single α-MHCcells with different morphologies (subpanels a, c, and e). Detection of α-cardiac actinin (subpanels b, d, and f) and connexion-43 (subpanels g and h) was performed using anti-cardiac actinin (1:400) and anti-connexin-43 (1:400). Secondary detection was performed with anti-mouse-IgG-AlexaFluor555 and anti-rabbit-Ig-AlexaFluor647. Hoechst dye was used to stain nuclei. Bars in panel e (subpanels a to f) are 50 μm; bar in panel e (subpanel g) is 20 μm; and bar in panel (subpanel h) is 7.5 μm

Hierarchical clustering of probe sets identified as downregulated in α-MHCcells

<p><b>Copyright information:</b></p><p>Taken from "Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes"</p><p>Genome Biology 2007;8(4):R56-R56.</p><p>Published online 11 Apr 2007</p><p>PMCID:PMC1896009.</p><p></p> Shown is a visualization of the hierarchical clustering of probe sets identified as downregulated in the α-MHCcells with an expression level at least twofold lower than in 15-day-old EBs and in α-MHC embryonic stem cells. Each probe set is represented by a single row of colored boxes; each array is represented by a single column. Rectangles corresponding to intermediately expressed probe sets are colored black, upregulated probe sets are indicated with red of increasing intensity, and weakly expressed probe sets with green of increasing intensity. The dendrogram on the left of the figure represents the similarity matrix of probe sets

Analysis of the vascular and haematopoietic cell gene markers in the BMP2cell population

RT-PCR analysis of the BMP2cells. Relative expression levels of the genes presented in (a) as obtained by Affymetrix analysis. The expression levels of each gene were normalized with its maximum level set as 100%. Each result is an average of three independent experiments (Additional data file 13). Immunuostainings with anti-pan cytokeratin over the period of time to show the presence of epithelial like cells, from left to right: one day after plating the BMP2cells (7 + 1 days in total); 8 days after plating with puromycin (7 + 8 days in total); 18 days after plating with puromycin (7 + 18 days in total); and 11 days after plating without puromycin (7 + 11 days in total).<p><b>Copyright information:</b></p><p>Taken from "Transcriptomic and phenotypic analysis of murine embryonic stem cell derived BMP2lineage cells: an insight into mesodermal patterning"</p><p>http://genomebiology.com/2007/8/9/R184</p><p>Genome Biology 2007;8(9):R184-R184.</p><p>Published online 4 Sep 2007</p><p>PMCID:PMC2375022.</p><p></p