Heat-map displaying gene expression profiles of Oxidative phosphorylation.

<p>The heat map represents the mRNA expression of Oxidative Phosphorylation genes downregulated in M1 condition. Gene coding for subunits of every components of the electron transport chain are represented. These genes tend to be upregulated in M2a 12–24 hours treatments.</p

Number of genes up- and down-regulated in polarized Mϕ at specific time points.

<p>Significantly differently expressed probes(p <0.05 moderated t test with Benjamini Hochberg)</p><p>Number of genes up- and down-regulated in polarized Mϕ at specific time points.</p

Enrichment map for M1 <i>vs</i> RM at 6, 12 and 24 hours.

<p>Panel A: the map displays the enriched pathways (p-value < 0.01, FDR < 20%), issued from GSEA, in M1 <i>vs</i> RM at 6 and 12 hours of culture. Enrichments were mapped to the inner node area and to the node borders, respectively. Panel B: the map displays the enriched pathways in M1 <i>vs</i> RM at 12 and 24 hours of culture. Enrichments were mapped to the inner node area and to the node borders, respectively. Red and blue dots represent up- and down-regulated gene-sets respectively, in M1 <i>vs</i>. RM. Edge thickness represents the fraction of common genes between pathways.</p

Validation of microarray data by quantitative pcr.

<p>mRNA expression level of <i>WNT5a</i>, <i>WNT5b</i>, <i>COX5a</i> and <i>GPD2</i> in RM, M1, M2a and M2c. axis: time course; Y axis: -ΔΔct.</p

Time-Series Clustering of M1 Microarray Gene Set Data Using STEM.

<p>The expression data were loaded onto STEM platform and distinct temporal expression profiles were generated, which differentiate between real and random patterns. Profiles are numbered from 0 to 19. Each box corresponds to a model expression profile. Significant expression profiles are highlighted in color. Clusters with similar colors show similar patterns. To all expression profiles a zero time point was added to serve the control value. Genes are assigned to the most closely matching profile by statistical analysis. The X-axis represents hours after stimuli addition and the Y-axis denotes fold-increase or decrease in expression in log₂ scale. Cluster were described with the main representative GO term.</p

Hierarchical clustering HCL.

<p>HCL analysis among resting (RM), classical (M1), alternative (M2a) and deactivate Mϕ (M2c) in the three point of time course (6, 12 and 24 h) on an ANOVA-filtered gene set. The red, yellow and green colors represent higher than average, close to average and lower than average gene expression respectively. X axis reports the type of activated Mϕ while genes are listed in the y axis.</p

Enrichment map for M2a <i>vs</i> RM at 12 and 24 hours.

<p>Panel A: the map displays the enriched pathways (p-value < 0.01, FDR < 20%), issued from GSEA, in M2a <i>vs</i> RM at 12 and 24 hours of culture. Enrichments were mapped to the inner node area and to the node borders, respectively. Red and Blue dots represent up- and down-regulated gene-sets respectively, in M2a <i>vs</i> RM. Edge thickness represents the fraction of common genes between pathways. Panel B: zoom in the cellular respiration cluster.</p

Clinical characteristics of the study population.

<p>Data expressed as mean±SD or N. Laboratory and diagnostic assays were from fasted blood samples.</p><p>HbA_1c = glycated hemoglobin;</p><p>HDL = high-denisty lipoprotein;</p><p>LDL = low-density lipoprotein;</p><p>GOT = glutamic-oxaloacetic transaminase;</p><p>GPT = glutamic-pyruvic transaminase;</p><p>BMI = body mass index;</p><p>SBP = systolic blood pressure;</p><p>DBP = diastolic blood pressure.</p><p>Cigarette smoking refers to current cigarette smoker.</p

Pioglitazone effect on EPC pro-inflammatory molecule expression.

<p>Effect of pioglitazone on PPARγ, adhesion molecule and TNFα expression in EPCs. PPARγ gene expression in early and late-outgrowth EPC exposed to pioglitazone (10 µM) and pioglitazone + GW9662 (1 µM) (A) Effect of pioglitazone in modulating ICAM-1 and VCAM-1 expression by early and late-outgrowth EPCs (cytofluorimetric analyses). Results are reported as delta % of MFI (mean fluorescence intensity) with vehicle condition (B). Effects of pioglitazone on TNFα gene (C) and protein (D) expression in early and late-outgrowth EPC. (*p<0.05 vs vehicle; **p<0.01 vs vehicle). Real time PCR data are expressed as −ΔΔCt and represent the relative gene expression of EPC cultured in the presence of pioglitazone 10 µM (with or without GW9662 1 µM) in relation to vehicle, normalized for the endogenous control GAPDH. Protein expression is measured with ELISA assay from culture supernatants. Results from five independent experiments performed in duplicate are shown (*p<0.05 vs vehicle) (Pio = pioglitazone).</p

Characterization of late-outgrowth EPCs.

<p>Morphology and phenotype of late-outgrowth EPC derived from human peripheral blood mononuclear cells. Colonies of late-outgrowth EPCs with a cobblestone-like morphology (A). Representative flow cytometry analysis of late-outgrowth EPCs (red) compared to early EPCs (purple) for the expression of CD31, KDR, CD146 and CD14 (B). Immunofluorescent staining of late-outgrowth EPCs for CD34, Ve-cad (vascular endothelial cadherin) and vWF (von Willebrand factor) (C), immunofluorescence staining of late-outgrowth EPC for DiI-LDL (red), lectin (green) and merge (bar = 50 µm) (D). eNOS and GAPDH gene expression in late EPCs (E) (bp = base pairs; eNOS = endothelial nitric oxide synthase; GAPDH = Glyceraldehyde 3-phosphate dehydrogenase).</p