The expression of the CD44 variant isoforms v3, v4, v4/5, v6, v7 and v7/8 in DLD-1 parental and <i>AKT</i>1/2 KO presented as the mean fraction (min-max value) of live cells from at least two flow cytometry experiments.

<p>The expression of CD44 standard variant was around 100%.</p><p>^*Mean fraction (min-max).</p

AKT expression in CD133/CD44 sorted cells.

<p>A) DLD-1 cells were sorted by flow cytometry and different populations with CD44^positive/CD133^negative (Q1), CD44^positive/CD133^positive (Q2), CD44^negativeCD133^negative (Q3), were collected. B) The sorted cells were further analyzed with western blot for total AKT, AKT1 or AKT2 and betaactin expression.</p

Protein and gene expression in DLD-1 parental, <i>AKT</i>1 KO, <i>AKT</i>2 KO, and <i>AKT</i>1/2 KO cells.

<p>A) Protein expression of CD44, CD133, phospho-FOXO, total FOX03a, phospho-GSK3β and total GSK3β from western blot analysis. B) Gene expression, up-regulation (+), down-regulation (−) or not changed (NC), of CD44, CD24, CD133, FOX01, FOX03, FOX04, GSK3β and LYN in DLD-1 <i>AKT</i>1 KO, <i>AKT</i>2 KO or <i>AKT</i>1/2 KO cells in comparison with DLD-1 parental cells.</p

Flow cytometry analysis of the expression of CD133, CD24 and CD44 in the colon cancer cell-line DLD-1 with its isogenic knock-out cell-lines of <i>AKT</i>1, <i>AKT</i>2 and <i>AKT</i>1/2.

<p>A) In the parental cells, approximately 10% of the cells were CD133 positive cells. However, in the <i>AKT</i>1 and <i>AKT</i>1/2 knock-outs, the CD133 positive cells were reduced to 0.3 and 0.1% respectively. This was not seen in the <i>AKT</i>2 knock-out cell-line, where 33% of the cells were positive for CD133. B) The mean fluorescent intensity of CD44 normalized to the DLD-1 parental cell-line increased to 150% in <i>AKT</i>1 KO, 160% in <i>AKT</i>2 KO and 300% in <i>AKT</i>1/2 KO cell-line. The error bars represent the standard deviation (SD) from at least two experiments. C) The percent of CD24 positive cells analyzed with two different CD24 antibodies from BD Biosciences and Miltenyi/MACS in flow cytometry. The standard deviations are from repeated experiments. D) Cell-cycle distribution in DLD-1 parental, <i>AKT</i>1 KO, <i>AKT</i>2 KO and <i>AKT</i>1/2 KO cells.</p

The log fold change of genes involved in important pathways in cancer in DLD-1 <i>AKT</i>2 KO versus <i>AKT</i>1 KO.

<p>Genes related to the WNT, Notch and cell adhesion molecules (CAMs) pathways were differently expressed.</p

Expression of CD133, CD44, CD24 and EGFR in three colon cancer cell-lines.

<p>A) HT-29, HCT116 and DLD-1. The expression patterns in the dotplots are from one representative flow cytometer experiment. The grid demonstrates the margin between high and low expression of the protein defined by isotype controls. B) The expression of CD24 positive cells in flow cytometry depend on the anti-CD24 antibody. The table shows the percent of CD24 positive cells using three different CD24 antibodies.</p

A template for writing radiotherapy protocols

<div><p></p><p><b>Background</b>. Well-specified and unambiguous treatment protocols are essential both for current practice and for the future development of radiation therapy. In order to provide assistance for writing good protocols, irrespective of treatment intention and complexity, up-to-date guidelines are highly desirable.</p><p><b>Methods</b>. We have analysed the radiotherapy work-flow, including clinical and physical aspects, such as preparatory imaging, treatment planning, delivery and evaluation, with the aim to outline a consistent framework covering the entire radiotherapy process.</p><p><b>Results</b>. Based on the analysis, a recipe-style template for specifying the description of the radiotherapy process has been designed. The template is written in a general format, which allows for modified phrasing, and should be customised for the specific clinical situation and diagnosis, as well as facility resources.</p><p><b>Conclusions</b>. The template can be used as a tool to ensure a consistent and comprehensive description of the radiotherapy section of clinical guidelines, care programmes and clinical trial protocols.</p></div

Associations between nuclear RBM3 expression and clinicopathological characteristics.

<p>ALP; alkaline phosphatase, CEA; carcinoembryonic antigen, KRAS; Kirsten rat sarcoma. (Regarding cytoplasmic expression see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0182512#pone.0182512.s001" target="_blank">S1 Table</a>).</p

Immunohistochemical images of RBM3 staining.

<p>Representing colorectal tumours with (A) negative, (B) weak, (C) moderate and (D-F) strong expression in a varying proportion of tumour cells. All images captured at X 20 magnification.</p

Treatment and treatment response.

<p>Divided according to high and low nuclear RBM3 expression, and positive and negative nuclear RBM3 expression. (Regarding cytoplasmic expression see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0182512#pone.0182512.s003" target="_blank">S3 Table</a>).</p