How should novelty be valued in science?

<p>Box plot analysis of serum concentrations of sRAGE (A), esRAGE (B), S100A9 (C) and HMGB1 (D) in patients with CTEPH (n = 26) and controls (n = 33). Independent Student’s t-test was used to compare groups. <i>RAGE</i> receptor for advanced glycation endproducts, <i>sRAGE</i> soluble RAGE, <i>esRAGE</i> endogenous secretory RAGE, <i>S100A9</i> member of S100 family of Ca+ binding proteins, <i>HMGB1</i> high mobility group box1, <i>CTEPH</i> chronic thromboembolic pulmonary hypertension.</p

Deletion of ATG7 in K14-expressing cells suppresses autophagy and leads to accumulation of p62 in the thymus.

<p>(<b>A</b>) Thymus lysates of the ATG7 f/f and ATG7 f/f K14-Cre, either carrying the GFP-LC3 transgene or not, were subjected to Western blot analysis for LC3 (upper panel) and p62 (lower panel). Bands corresponding to LC3-I and LC3-II (indicative of active autophagy) are marked by arrows. Note the presence of LC3-II bands in ATG7 f/f thymi and the accumulation of LC3-I as well as p62 in ATG7 f/f K14-Cre mice. The position of an unspecific band, that shows equal loading of the lanes, is marked by an asterisk. (<b>B</b>) Immunofluorescence analysis of p62 expression (red) in the thymus. Note the accumulation of p62 in the characteristically shaped epithelial cells of ATG7 f/f K14-Cre mice. Nuclei are labeled with Hoechst 33258 (blue). The complete field of view under 200-fold magnification is shown in each panel.</p

Tissue inflammation is not increased by suppression of autophagy in the thymic epithelium.

<p>The severity of inflammation in organs of mice expressing ATG7 (ATG7 f/f) or lacking ATG7 in the thymic epithelium was scored using H&E-stained sections. Scores 0 (green portion of the bars), 1 (yellow) and 2 (orange) mean no inflammation, mild and moderate inflammation (respectively). The bars show the percentage of mice with the respective score. The number of mice of each group is shown below the bars.</p

Deletion of ATG7 in K14-expressing cells suppresses autophagy in the thymic epithelium of GFP-LC3 transgenic mice.

<p>Thymi from GFP-LC3 ATG7 f/f and ATG7 f/f K14-Cre were cryo-sectioned and immunolabeled with anti-keratin 8 (K8) (<b>A</b>) and anti-K5 antibodies (red) (<b>B</b>). The immunolabeling and the green fluorescence of the GFP-LC3 fusion protein were viewed under a confocal laser scanning microscope. Regions of the thymus cortex (<b>A</b>) and medulla (<b>B</b>) are shown. Arrows in A point to GFP-LC3 puncta corresponding to labeled autophagosomes. Differently shaped arrows point to green fluorescent dots of uncertain identity that were detected at low frequency in the medulla (<b>B</b>). Arrowheads in <b>A</b> and <b>B</b> indicate diffuse cytosolic accumulation of GFP-LC3 in the epithelial cells of the cortex (<b>A</b>) and the medulla (<b>B</b>). Bar, 10 µm.</p

T cell differentiation into CD4+ and CD8+ cells and activation of CD4+ cells are not compromised in ATG7 f/f K14-Cre mice.

<p>Lymphocytes were prepared from the thymus (<b>A</b>, <b>B</b>), lymph nodes (<b>C</b>, <b>D</b>) and spleen (<b>E</b>, <b>F</b>) and subjected to FACS with antibodies against CD3, CD4, and CD8 (<b>A</b>, <b>C</b>, <b>E</b>) as well as with antibodies against CD4 and CD69 (<b>B</b>, <b>D</b>, <b>F</b>). Statistical analysis using the t-test showed that there were only non-significant (n.s.) differences between the genotypes. The results of one of two experiments with similar results are shown.</p

The weights of the body, thymus and spleen of ATG7 f/f K14-Cre mice are normal.

<p>The weights of the total body (<b>A</b>), the thymus (<b>B</b>) and the spleen (<b>C</b>) were determined for male mice in the age range from 6 to 12 months. Note that data for female mice are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038933#pone.0038933.s001" target="_blank">figure S1</a>. The weight data of mice expressing ATG7 (ATG7 f/f) (n = 14) or lacking ATG7 (ATG7 f/f K14-Cre) (n = 11) in the thymic epithelium were compared in absolute numbers or relative to the body weight (<b>D</b>, <b>E</b>). Statistical analysis using the t-test showed that there were only non-significant (n.s.) differences between the genotypes.</p

SEC^PBMC leads to enhanced wound closure and re-epithelialization.

<p>(<b>A</b>) Wound areas were measured during the first 3 days after wounding. Treatment with SEC^PBMC significantly enhanced wound closure. Error bars represent one standard deviation calculated from 15 animals for each set of values (*: p<0.05). (<b>B</b>) Representative photographs from mouse wounds (n = 15 from each group) immediately after wounding and at day 7 after wounding are shown. (<b>C</b>) H&E staining of wounds treated with medium or SEC^PBMC 7 days after wounding is shown. While medium treated wounds still show a thick crust and little re-epithelialization, SEC^PBMC treated wounds are fully re-epithelialized. C  =  crust, E  =  newly formed epidermis, G  =  granulation tissue. Scale bars: 100 µm. One representative animal of 15 is shown.</p

Apoptotic MNC-secretomes in experimental stroke

<p>Mixed Model Analysis (SAS output)</p> <p>The data were analyzed using linear mixed models for the neuroscore on treatment group and time-point with the factor animal included as a random effect. The MIXED procedure in SAS 9.3 was used to perform the calculations. The raw output contains information on the model specifications, the estimated error variance and random effects variance, the estimated regression coefficients, the covariance structure of the model coefficients and type III F-tests for the hypotheses of no effect of either fixed effect or their interactions. An interaction plot was drawn using the GLM procedure. This plot shows the individual observations and their sample mean values in each group and for each time-point. The group labels 0,1,2 and 3 in the raw output refer to the treatment group in setting 1, the control group in setting 1, the treatment group in setting 2 and the control group in setting 2, respectively.</p> <p>Original Western blots to Figure 5 </p> <p>Expression of proteins involved in cytoprotective pathways in human Astrocytes and Schwann Cells </p> <p>Astrocytes (page 1) or Schwann Cells (page 2) were stimulated with hMNCapo sec, control medium (served as control to treatment) or positive control (control to the measured protein). Original blots for all measured proteins are given in this raw data set (pages 1 and 2). For each blot, lanes (1), (2), and (3) correspond to the groups medium control [(1)=control to treatment], human apoptotic MNC-secretomes [(2)=treatment] and positive control [(3)=recombinant protein].<br>Bands in each blot are shown for phosphorylated CREB, total-CREB, phosphorylated Erk1/2, total-Erk 1/2, phosphorylated HSP27, total-HSP27, phosphorylated cJun, total-cJun, phosphorylated Akt, and total-Akt. The molecular weight (kDa) for each protein can be seen under each blot.<br>Ponceau staining was used as loading control for each group (1), (2), and (3) and suggest equal loading.</p> <p>Original Western blots to Figure 6 </p> <p>Expression of Phosphorylated CREB in Astrocytes and Neurons after stimulation with the active compound hMNCapo sec or control medium: </p> <p>Cultured human Astrocytes and Neurons were incubated with hMNCapo sec or control (cell culture-) medium at indicated concentrations. Original blots can be seen here.<br>Ponceau staining shows equal loading.</p> <p> </p

Five samples, which were negative and 5 samples, which showed positive immunostaining, were used for PCR.

<p>The MLK1 Merkel cell carcinoma cell line was used as positive control for the establishment of the MCPyV PCR (Fig 2).</p

List of Antibodies.

<p>Company addresses:</p>1<p>Bedford, MA, USA; ²Littleton, CO, USA; ³Cambridge, UK; ⁴NEB: New England Biolabs, Beverly, MA, USA; ⁵Turku Finland; ⁶Buckinghamshire, UK; ⁷Rockford, IL, USA; ⁸Eugene, OR, USA. n.d.: not done.</p