Plasmid- and chromosomal genes involved in Corrin and Adenosyl cobalaimin (B12) uptake, salvage and biosynthesis in <i>Azotobacter chroococcum</i> NCIMB 8003.

<p>A. The region of 16,967 bp from bp 64,013 to 80,225 in plasmid pAcX50f containing a contiguous cluster of genes for corrin synthesis. B. The regions of 12,882 from bp 1,777,467 to 1,789,349; 740,868 to 741,638 and 3,736,258 to 3,738,156 in the chromosome potentially containing genes for corrin and B12 uptake and salvage and the late steps in the synthesis of adenosyl cobalamin.</p

Ac-8003 and Av-DJ Genomes Compared.

<p>Ac-8003 and Av-DJ Genomes Compared.</p

The Methylome of <i>Azotobacter chroococcum</i> NCIMB 8003.

<p>^m6A = <i>N</i>⁶-methyladenine, ^m4C = <i>N</i>^<i>4</i>-methylcytosine; W = A or T, S = C or G</p><p>The Methylome of <i>Azotobacter chroococcum</i> NCIMB 8003.</p

Physical and genetic map of plasmid pAcX50c from A. chroococcum NCIB 8003.

<p>The map shows the arrangement of deduced genes in the sequence of the potential retro-conjugative plasmid pACX50c. The two clusters of contiguous <i>tra</i> and <i>trb</i> conjugation genes are shown in green and dark blue respectively. Genes for other functions are indicated as follows: replication initiation (<i>trfA</i>: red); partitioning and maintenance (pink), Type I RMS system (<i>Ach</i>III) (yellow); transposases (brown); DNA binding and repair (grey); unidentified or conserved unidentified genes (white).</p

Comparison of the gene contents in the chromosomes of <i>A</i>. <i>chroococcum</i> NCIMB 8003 and <i>A</i>.<i>vinelandii</i> DJ (ATCC BAA-1303).

<p>Genes present in both strains are shown in red (“core genes”) whilst the genes present in only that strain (“accessory genes”) are shown in blue. The genome dimensions are marked in bp.</p

Synteny between the chromosomes of <i>A</i>. <i>chroococcum</i> NCIMB 8003 (ordinate) and <i>A</i>.<i>vinelandii</i> DJ (ATCC BAA-1303).

<p>The nucleotide sequences of the chromosomes of <i>A</i>. <i>chroococcum</i> NCIMB 8003 (ordinate) and <i>A</i>.<i>vinelandii</i> DJ (ATCC BAA-1303) (abscissa) were aligned using BLASTn. The red and blue diagonal lines show the slopes that would be obtained were each chromosome aligned against itself: red, <i>A</i>. <i>chroococcum</i>; blue, <i>A</i>.<i>vinelandii</i>.</p

MOESM2 of Obesity inhibits the osteogenic differentiation of human adipose-derived stem cells

Additional file 2: Figure S2. Estradiol enhanced osteogenic differentiation of obASCs. lnASCs and obASCs were seeded on PLGA scaffolds and induced with CDS-ODM supplemented with 10 Nm estradiol. Scaffolds stained with Alizarin Red are shown. Scale bar represents 1 mm

MOESM1 of Obesity inhibits the osteogenic differentiation of human adipose-derived stem cells

Additional file 1: Figure S1. Estradiol restores the osteogenic differentiation capacity of obASCs. (A) lnASCs (n=6 donors) and obASCs (n=6 donors) were cultured in CDS-ODM supplemented with estradiol. After 14 days, cells were stained with Alizarin Red. Scale bar represents 200 μm. (B) To quantify the amount of Alizarin Red staining, stains were eluted with CPC and optical density was read. Bars, ± SEM. *, P < 0.05; **, P < 0.01 between lnASCs and obASCs

LPA gradients across melanomas <i>in vivo</i>.

<p>(A) TYR::CreER^T2BRAF^V600E/+PTEN^lox/+ mice, a genetically appropriate melanoma model, were treated with tamoxifen as described in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966-Garcia1" target="_blank">[56]</a>, grown until melanomas spontaneously developed. Dashed box shows the region used for the samples shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio-1001966-g001" target="_blank">Figure 1C</a>. (B) Haematoxylin and eosin-stained biopsies of murine melanomas demonstrating the dispersal of cells from a representative tumour from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio-1001966-g005" target="_blank">Figure 5A</a>, with cells spreading directly away from the tumour. Upper image 2.5× magnification; lower image 20× magnification from dashed box above, showing melanoma cells invading toward the muscle layer (D, dermis; M, muscle layer). (C) Biopsies from mouse melanomas. Several sites in a linear distribution were biopsied using a 6 mm punch biopsy tool within 5 minutes of the mouse being sacrificed and immediately frozen in liquid nitrogen. The positions of biopsies used for LPA measurement are indicated (too few distant samples were obtained for a significant measurement). Bar shows 5 mm. (D) LPA concentration gradients across the margin of a melanoma. Four melanomas were sampled at three sites in a line as shown in (A) (A, tumour body; B, tumour edge; C, skin surrounding tumour). Total LPA per mg tissue was quantified by mass spectrometry after weighing the tissue specimens and extracting the LPA. Outward-directed gradients of LPA were found across the margin of all the melanomas tested. Bars show SEM. (E) Analysis of LPA subspecies. 18∶2-LPA, 20∶4-LPA and 22∶6-LPA show a clearer gradient than 16∶0-LPA, which is though to be less active as a signalling molecule.</p

Growth factors potentiate LPA chemotaxis.

<p>(A) Growth factors enhance cells' response to LPA gradients. Figure shows plots of the WM239A paths chemotaxing in gradients of LPA, LPA+EGF+PDGF, and conflicting gradients of LPA versus EGF+PDGF. (B) Chemotactic indices of cells in (A) and other conditions. Growth factor gradients if anything increase the efficiency of LPA chemotaxis, even when applied in a gradient in the opposite direction. Bars show SEM.</p