Additional file 2: Figure S2. of Metallothionein regulates intracellular zinc signaling during CD4+ T cell activation

T helper cell inducing conditions have an effect on the intracellular [Zn2+] of CD4+ T cells during primary activation and differentiation. Splenocytes from MT knockout (MT-/-) (n = 3–4) or wildtype controls (MT+/+) (n = 3–4) were stimulated with anti-CD3 and anti-CD28 for 6 days in the presence of no added cytokines (Th0), IL-12 + IL-2 + anti-IL-4 (Th1), or IL-27 (Tr1) to promote differentiation and expansion of T helper cell populations. Intracellular [Zn2+] in the CD4+ T cell population was measured for each inducing condition and MT genotype at +6 days post stimulation. (PDF 90 kb

Additional file 4: Figure S4. of Metallothionein regulates intracellular zinc signaling during CD4+ T cell activation

Addition of ZnSO4 [20μM] to cell culture media for 24 h increases intracellular [Zn2+] in CD4+ Tr1 cells. Tr1 cells from MT knockout (MT-/-) (n = 6) or wildtype control (MT+/+) mice (n = 6) were incubated in media + ZnSO4 [20μM] or media alone for 24 h. Tr1 cells were loaded with fluozin-3 AM and the intracellular [Zn2+] was measured by flow cytometry. (PDF 98 kb

Additional file 1: Figure S1. of Metallothionein regulates intracellular zinc signaling during CD4+ T cell activation

Increasing extracellular [Zn2+] does not affect intracellular Zn2+ release following exposure to ROS. Tr1 cells from metallothionein wildtype control mice were exposed to 5ÎźM H2O2 in media (circle) or media with 20ÎźM ZnSO4 added (square). Baseline Fluozin-3 fluorescence was established for 30 s followed by a 7 min exposure to H2O2. (PDF 109 kb

Degradation of Bunker C Fuel Oil by White-Rot Fungi in Sawdust Cultures Suggests Potential Applications in Bioremediation

<div><p>Fungal lignocellulolytic enzymes are promising agents for oxidizing pollutants. This study investigated degradation of Number 6 “Bunker C” fuel oil compounds by the white-rot fungi <i>Irpex lacteus</i>, <i>Trichaptum biforme</i>, <i>Phlebia radiata</i>, <i>Trametes versicolor</i>, and <i>Pleurotus ostreatus</i> (Basidiomycota, Agaricomycetes). Averaging across all studied species, 98.1%, 48.6%, and 76.4% of the initial Bunker C C10 alkane, C14 alkane, and phenanthrene, respectively were degraded after 180 days of fungal growth on pine media. This study also investigated whether Bunker C oil induces changes in gene expression in the white-rot fungus <i>Punctularia strigosozonata</i>, for which a complete reference genome is available. After 20 days of growth, a monokaryon <i>P</i>. <i>strigosozonata</i> strain degraded 99% of the initial C10 alkane in both pine and aspen media but did not affect the amounts of the C14 alkane or phenanthrene. Differential gene expression analysis identified 119 genes with ≥ log₂(2-fold) greater expression in one or more treatment comparisons. Six genes were significantly upregulated in media containing oil; these genes included three enzymes with potential roles in xenobiotic biotransformation. Carbohydrate metabolism genes showing differential expression significantly accumulated transcripts on aspen vs. pine substrates, perhaps reflecting white-rot adaptations to growth on hardwood substrates. The mechanisms by which <i>P</i>. <i>strigosozonata</i> may degrade complex oil compounds remain obscure, but degradation results of the 180-day cultures suggest that diverse white-rot fungi have promise for bioremediation of petroleum fuels.</p></div

Biplot of principal components (PC) axes PC1 and PC2 derived from a Principal Components Analysis (PCA) of mapped paired-end Illumina RNA-Seq reads from 20 day-old <i>Punctularia strigosozonata</i> cultures in media treatments (aspen, aspen with Bunker C oil, pine, and pine with Bunker C oil).

<p>Ordination was completed in the R package DESeq.</p

Hydrocarbon degradation by <i>Trichaptum biforme</i>.

<p>GC-MS chromatograms of (a) alkane and (b) phenanthrene degradation by <i>T</i>. <i>biforme</i> measured after 180 days of growth in pine media with Bunker C oil. Black lines = <i>T</i>. <i>biforme</i> profiles; blue lines = Bunker C oil profiles.</p

<i>Punctularia strigosozonata</i> transcripts with predicted protein functions expressed ≥ log₂(2-fold) (adjusted p < 0.01) in comparisons of 20-day growth on aspen and pine media with and without Bunker C oil.

<p>Positive log₂ fold changes indicate transcript accumulation in the first treatment while negative log₂ fold changes indicate transcript accumulation in the second treatment.</p><p><i>Punctularia strigosozonata</i> transcripts with predicted protein functions expressed ≥ log₂(2-fold) (adjusted p < 0.01) in comparisons of 20-day growth on aspen and pine media with and without Bunker C oil.</p

Degradation (%) of phenanthrene, a C14 alkane, and a C10 alkane in Bunker C oil by white-rot fungi.

<p>Degradation percentages were calculated using <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130381#pone.0130381.e001" target="_blank">Eq 1</a>.</p><p>Degradation (%) of phenanthrene, a C14 alkane, and a C10 alkane in Bunker C oil by white-rot fungi.</p

Illumina RNA-Seq read counts and percentages of reads mapped to the <i>Punctularia strigosozonata</i> genome and transcriptome.

<p>Illumina RNA-Seq read counts and percentages of reads mapped to the <i>Punctularia strigosozonata</i> genome and transcriptome.</p