<i>TGM1</i>, <i>TGM3</i> and <i>TGM5</i> gene expression in the skin of AD patients and healthy controls.

<p>TGM transcript levels (A, E and I) of healthy controls (HC, n = 10) non lesional skin of AD patients (NL, n = 7) and lesional skin from AD patients (L, n = 10). Horizontal bars represent median values in each group and data is presented on a logaritmic scale. For IHC analysis of the TG protein expression, skin sections from nine AD patients and ten healthy controls were stained. Representative staining from one healthy control and one patient is shown in the figure for TG1 (B–D), TG3 (F–H) and TG5 (J–L) expression. Scale bar represents 50 µm.</p

Cell morphology, lineage tracing, FACS sort and proliferation rates of mucoepidermoid tumor before and after sorting.

<p>Mucoepidermoid tumor cells at passage 0, Scale bar: 100 µm (a), BM-MSCs at passage 6, Scale bar: 100 µm (b), a representation of the merged images of MUC 1 and MAML2 before, Scale bar: 20 µm (c, d) and after, Scale bar: 50 µm (h, i) sort, flow cytometry sorting for a panel of mesenchymal stromal cell markers (e), post-sorting of MEi cells after 24 hours recovery, Scale bar: 500 µm (f) and a representative image of expanded post-sorted p12+5 MEi cells, Scale bar: 500 µm (g), cell proliferation rates and doubling times of unsorted and sorted cells (black and red respectively) <i>versus</i> BM-MSC (blue) (j). Colored circle indicates the raw datasets for the number of cells in each well; crosses and error bars indicate means and standard deviations.</p

Histological and pathological analyses of a mucoepidermoid tumor.

<p>Haematoxylin and Eosin staining provides visualization of the excised tracheal mucoepidermoid tumor biopsy (a), periodic acid-Schiff stain identifies mucins and glycoproteins in magenta (b), Mucin 1, Muc-1 identifies mucin-producing cells (c), Cytokeratin marker, Ck-MNF116 shows epithelial staining (d), carcino-embryonic antigen, CEA is an oncofetal antigen expressed by some tumors but not in normal adult tissues (e), androgen receptor, AR is a predictive marker for choice of therapy (f), muscle specific actin, MSA identifies the stromal component (g), and Ki-67 is a cell proliferation marker used for quantifying the proliferative index of the tumor (h). Scale bar: 100 µm.</p

Gene expression profiles and karyotyping of MEi cells.

<p>Two-dimensional cluster plots for the classification of samples based on the first two principal components. Tumor 1, 2, 3, 4 are MEi cells and MSC 1, 2 are BM-MSCs (a), Each row represents a gene and each column represents a sample. The expression level of each gene in a single sample is relative to its median abundance across all samples and is depicted according to a color scale shown on the right. Red and green indicate expression levels respectively above and below the median. (b), a representative image of tetraploidy and normal karyotype (c) at passage 12+3.</p

<i>In vitro</i> differentiation of MEi (MSC-like mucoepidermoid tumor) cells to endoderm and ectoderm lineages.

<p>A representative image of endoderm differentiation of MEi cells (a–c) and BM-MSCs (d–f) were stained positive for GATA 6, and ectoderm differentiation of MEi cells (g–i) and BM-MSCs (j–l) were positive for β-III tubulin. Scale bar: 50 µm.</p

Spheroid cultures and flow cytometry analyses of spheroid outgrowths.

<p>Day 10 spheroids in low adhesion culture plates (a), day 10 trachea spheroids plated onto culture plates after 24 hours (b), outgrowths from spheroid cultures (c). Flow cytometry analyses of spheroid outgrowths after 25 days <i>in vitro</i>. Scale bar: 100 µm. Cell morphology before (d) and after (f) trypsinization. Scale bar: 200 µm. Flow cytometry analyses for a panel of mesenchymal stromal cell markers before (e) and after (g) trypsinization.</p

KEGG pathways significantly enriched by genes differentially expressed between MEi cells and BM-MSCs.

<p>Pathways were identified by DAVID Functional Annotation and ranked by P-value with a cutoff of 0.01. Counts and percentages refer to the number and percentage of genes from the input list that fit into a given KEGG pathway. Fold enrichment is the magnitude of enrichment for each KEGG pathway compared with the entire gene list in the Affymetrix Human Gene 1.1 ST Array that serves as the reference.</p><p>KEGG pathways significantly enriched by genes differentially expressed between MEi cells and BM-MSCs.</p

Differentiation of MEi (MSC-like mucoepidermoid tumor) cells to mesenchymal trilineage.

<p>Directed differentiation with growth factors (a–c): towards osteoblast phenotype (alizarin Red staining) (a), towards adipocyte phenotype (Oil red staining) (b), towards chondrocyte phenotype (toluidine blue staining) (c); Spontaneous differentiation without growth factor (d–f). alizarin S staining (d) oil red staining (e), toluidine blue staining (f). Scale bars: 100 µm. MicroRNA analyses of miR-34, miR 449a, b, c for BM-MSC, unsorted, sorted and spontaneous differentiation of tracheal tumor cells (g).</p

Partial list of differentially expressed genes from the KEGG annotation: Pathways in cancer.

<p>Log₂FC is the base 2 logarithm of the fold change, with negative values indicating that the gene is more highly expressed in the tumor samples.</p><p>Partial list of differentially expressed genes from the KEGG annotation: Pathways in cancer.</p