MOESM2 of Influence of different wastewater treatment technologies on genotoxicity and dioxin-like toxicity in effluent-exposed fish

Additional file 2.  All data

MOESM1 of Influence of different wastewater treatment technologies on genotoxicity and dioxin-like toxicity in effluent-exposed fish

Additional file 1: Semi-quantitative histolopathological assessment of liver, kidney and gill samples of fish exposed up- and downstream of WWTP B. Samples were classified into five categories described in detail by Wilhelm et al. [17] according to the symptoms displayed. Shortly, class 1 includes samples in control state, class 2 has been assigned to tissue samples displaying slight reactions, class 3 to samples with pronounced reactions, class 4 describes samples expressing beginning destructive alterations and class 5 has been assigned to liver samples with severe cellular destruction. Asterisks and horizontal lines indicate significant differences between two datasets according to likelihood ratio chi-square tests with subsequent Holm correction

Quantification of Prohormone Processing Components in Human Dense Core Secretory Vesicles (DCSV).

<p>The NASF values of the relative abundances for prohormone proteins and prohormone processing enzymes are listed as NASF ± S.D. (×10³). The prohormone processing enzymes represent two protease pathways consisting of (1) the subtilisin-like prohormone convertases (PC1/3 and PC2) with their endogenous regulators (proSAAS and 7B2 regulators, respectively), combined with carboxypeptidase E (CPE), and the (2) cysteine protease cathepsin L pathway with aminopeptidase B (AP-B).</p

Functional categories of human DCSV soluble and membrane proteins.

<p>Pie charts illustrate the relative portion of proteins in each functional category for the soluble (panel A) and membrane (panel B) fractions of human DCSV. Each functional category, with name and percent of the total number of DCSV proteins, of the pie chart is shown as a distinct color. The proteins comprising each functional category are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone.0041134.s006" target="_blank">Table S3</a>, and proteomics identification of soluble and membrane proteins of human DCSV are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone.0041134.s004" target="_blank">Tables S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone.0041134.s005" target="_blank">S2</a>.</p

Cytoscape systems biology analyses of the human DCSV proteome.

<p>Components of the DCSV proteomics data were analyzed by the Cytoscape systems biology program for predicting protein interaction networks. The functional protein categories are illustrated on the right hand side. Based on quantitative NASF data of the proteins, individual proteins are indicated as predominantly soluble (green circles), predominantly membrane (red circles), or present in both soluble and membrane at similar levels (yellow) (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone-0041134-g003" target="_blank">Figure 3</a>). These color-coded protein symbols are those which were quantitated by NSAF. Proteins illustrated by grey circles are those which were identified, but not quantitated since they did not meet the criteria for quantitation in at least 3 out of 4 nano-LC-MS/MS runs.</p

Quantification of protein categories of human dense core secretory vesicles.

<p>Relative quantification of proteins in the main functional categories were assessed by normalized spectral abundance factors (NSAF), as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone.0041134.s001" target="_blank">Experimental Procedures S1</a>. Bar graphs illustrate average NSAF (sum) for each protein group with s.e.m. NSAF values for human DCSV proteins are provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041134#pone.0041134.s007" target="_blank">Table S4</a>.</p

Number of Proteins Identified and Quantitated Among Functional Categories of Human Dense Core Secretory Vesicles (DCSV).

<p>The number of distinct proteins identified and quantitated (in parentheses) in each of the functional categories are indicated for soluble and membrane proteins of the human DCSV.</p

Systems biology analyses distinguishes protein kinase A (PKA) and protein kinase C (PKC) pathways in human DCSV.

<p>Human DCSV proteins interacting with PKA or PKC are illustrated in green or red circles, respectively. Proteins interacting with both PKA and PKC are shown in yellow.</p

Differential regulation of neuropeptides, catecholamines, and β-amyloid secretion by activation of PKA and PKC in neuronal-like chromaffin cells.

<p>Adrenal medullary chromaffin cells in primary culture (bovine) were treated with forskolin that activates protein kinase A (PKA) that activates PKA via adenylyl cyclase stimulation and cAMP production, or with PMA (phorbol myristate acetate) that directly activates PKC. In time-course studies for treatment with forskolin or PMA for 15 minutes to 6 hours, the media was collected for measurement of secreted (Met)enkephalin and galanin neuropeptides (panels a and b, respectively), beta-amyloid peptide (Aβ(1–40), panel c), and the catecholamines dopamine, norepinephrine, and epinephrine (panels d, e, and f, respectively). Data for Control untreated cells (○), forskolin-treated cells (•), and PMA-treated cells (X) are plotted. Data for each time point represents the mean ± s.e.m. (n = 6); error bars are illustrated, and they are often smaller than the symbol.</p