Additional file 1 of Analysis and prediction of interactions between transmembrane and non-transmembrane proteins

Supplementary Material 1

Overexpression of neogenin in SHG-44 cell line.

<p>(A) Western blot results showed neogenin expression in the blank control which was untreated (Con), the negative control which was transfected by empty vector (Vec) and the over-expressive group which was transfected by neogenin expression plasmid (Neo) in SHG-44 cell line at 48 hours after transfection; (B-D) Status of cells in the blank, negative and over-expressive group of neogenin respectively at 48 h after transfection. (E) Apoptotic distribution maps of cells in the blank, negative and over-expressive group were drew by flow cytometry assay respectively. (F) Average apoptotic rate of cells in the blanck, negative and over-expressive group in the flow cytometry assay, One-Way ANOVA, n = 3, **<i>p</i><0.01, error bars indicate standard error means.</p

The 69 patients’ classified statistic according to clinical and pathological features (January, 2006 – January, 2011).

<p>IOD: integral optical density;</p>*<p>: independent <i>t</i>-test,</p>†<p>: One-Way ANOVA. All tumor sizes were measured by CT or MRI except the data from visual inspection in the operation (details in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038074#pone.0038074.s002" target="_blank">Table S2</a>),</p>‡<p>: n = 57. IOD values were supplied as Mean±Standard Deviation.</p

Clinicopathologic features and relative neogenin level profile in 13 paired surrounding and glioma tissues (January – November, 2011).

<p>SD: standard deviation;</p>*<p>: paired <i>t</i>-test.</p

Immunohistochemical analysis of neogenin in primary gliomas.

<p>(A–D) Representative immunohistochemical pictures of grade I-IV glioma respectively, (A’–D’) Magnification of the red squares in (A–D), (A–D) 200× magnification; (A’–D’) 400× magnification. (E) Scatter diagram of neogenin expression in 69 primary gliomas, blue bars are the means of four grades, n = 69, One-Way ANOVA was used in statistical analysis. (F) Histogram of mean neogenin expression in low-grade gliomas and high-grade gliomas, statistical analysis was performed with independent <i>t</i>-test. *<i>p</i><0.05, **<i>p</i><0.01, error bars indicate standard error means.</p

Multivariate hazard ratios of progression to high-grade gliomas in 69 primary glioma patients (January, 2006 – January, 2011).

<p>Multivariate hazard ratios and 95% Confidence Intervals were obtained by using Latency as time-scale and High-grade glioma as Failure event. ^‡: divided by Median of neogenin IODs listed in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038074#pone.0038074.s002" target="_blank">Table S2</a>.</p

Kaplan-Meier survival curves for overall progression to high-grade glioma.

<p>Failure event for computation of this curve was diagnosed as the high-grade glioma. Higher neogenin patients are marked in green color, lower neogenin patients are marked in blue color.</p

Primary and recurrent clinicopathologic features of 16 patients (January, 2001 – January, 2011).

<p>IOD: integral optical density;</p><p>IOD values were supplied as Mean±Standard Deviation.</p>*<p>: paired <i>t</i>-test;</p>†<p>: undefined grade gliomas.</p

Expression of neogenin in 16 paired primary and recurrent glioma sections.

<p>(A-B) Representative immunohistochemical pictures (200× magnification): (A) primary glioma, (B) matching recurrent glioma. (C) statistical graph of mean neogenin expression in primary and recurrent gliomas. Paired t-test, n = 16, * <i>p</i><0.05, error bars indicate standard error means.</p

Clinicopathologic features and the methylation of neogenin in 4 non-neoplastic brain tissues and 29 glioma tissues (January – November, 2011).

<p>NB: non-neoplastic brains;</p>*<p>: Chi-square test;</p>†<p>: the compare between the sum of NB and low-grade gliomas and high-grade gliomas.</p