Environmental Themes as a Way to Evaluate Pupils' Civil Competence in Chemistry Education at Primary Schools

Results of proteomic relative quantifiction of nuclear extracts from ES and ELA cells; Genes, unrelated to chromatin remodelers, with similar regulation of mRNAs in Fig. 4b–d. (XLS 24 kb

CSB ablation induced apoptosis is mediated by increased endoplasmic reticulum stress response - Table 1

<p>CSB ablation induced apoptosis is mediated by increased endoplasmic reticulum stress response</p> - Table

Src kinase regulates the RACK1/ZBP1 complex and the Y246 of RACK1 is critical for the binding with ZBP1.

<p><b>A</b>, Endogenous RACK1 interacts with Flag-ZBP1 in an anti-Flag immunoprecipitation assay from total lysate of neuroblastoma Flag-ZBP1-transfected cells. Western blotting for endogenous RACK1 and Flag-ZBP1 on proteins eluted from anti-Flag immunoprecipitation assay. Flag transfected cells were used as negative control. Input represents 5% of total lysate. <b>B</b> and <b>C</b>, Src activity regulates the RACK1-ZBP1 complex formation. <b>B</b>, db-cAMP or PACAP treatments of Flag-ZBP1 transfected cells reduced the binding between RACK1 and Flag-ZBP1, whereas PP2 restored the binding, as in untreated cells. <b>C</b> BDNF treatments increased the binding of Flag-ZBP1 to RACK1 in Flag-ZBP1 transfected cells. Src inhibition by Src inhibitor PP2 reduced the RACK1-ZBP1 complex stimulated by BDNF. The density value of immnoprecipitated RACK1 is normalized to that of immunoprecipitated Flag-ZBP1 and summarized in both graphics. Data are graphed as means plus S.D. <b>D</b>, The Src binding and phosphorylation site (Y246) of RACK1 is critical for complex formation. Flag-ZBP1 protein co-immunoprecipitated with GFP-RACK1_wt and GFP-RACK1_Y246F in immunopreciptation assays using anti-GFP antibody, but in the presence of GFP-RACK1_Y246F the binding was reduced. The figures are representative of three independent experiments. The density value of co-immnoprecipitated Flag-ZBP1 is normalized to that of immnoprecipitated GFP-RACK1_wt or GFP-RACK1_Y246F and summarized in both graphic. Data are graphed as means plus S.D.</p

Filtered list of genes which showed a fold change > 2 (ASO vs SO) in the microarray analysis at 6h, and belonged to the top GO terms listed in Table 1.

<p>Filtered list of genes which showed a fold change > 2 (ASO vs SO) in the microarray analysis at 6h, and belonged to the top GO terms listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172399#pone.0172399.t001" target="_blank">Table 1</a>.</p

Molecular mechanism of RACK1/ZBP1 complex regulating the release and translation of β-actin mRNA.

<p>A, Schematic figure showing the interaction of RACK1 with the β-actin mRNA/ZBP1 complex through its Src binding site (Y246) on 40S ribosome subunit of RNPs (<i>left</i>). In presence of the mutation Y246F (<i>right</i>), RACK1 on ribosomes fails to recruit the β-actin mRNA/ZBP1 complex and Src. This impairment blocks the release of β-actin mRNA from ZBP1 and its translation. B, Src can be activated by BDNF or by PACAP treatments. In BDNF stimulation, Src, activated through TrkB, may directly bind, and phosphorylate, both free or ribosomal bound RACK1 and free β-actin mRNA/ZBP1 complex. Next, RACK1 and β-Actin mRNA/ZBP1 associate on ribosomes and the β-actin mRNA is released to be translated. Instead, PACAP-activated Src stimulates PKA kinase which in turn may induce the dissociation of the β-actin mRNA/ZBP1-RACK1 complex by phosphorylating ZBP1.</p

CSB ablation induced apoptosis is mediated by increased endoplasmic reticulum stress response

<div><p>The DNA repair protein Cockayne syndrome group B (CSB) has been recently identified as a promising anticancer target. Suppression, by antisense technology, of this protein causes devastating effects on tumor cells viability, through a massive induction of apoptosis, while being non-toxic to non-transformed cells. To gain insights into the mechanisms underlying the pro-apoptotic effects observed after CSB ablation, global gene expression patterns were determined, to identify genes that were significantly differentially regulated as a function of CSB expression. Our findings revealed that response to endoplasmic reticulum stress and response to unfolded proteins were ranked top amongst the cellular processes affected by CSB suppression. The major components of the endoplasmic reticulum stress-mediated apoptosis pathway, including pro-apoptotic factors downstream of the ATF3-CHOP cascade, were dramatically up-regulated. Altogether our findings add new pieces to the understanding of CSB mechanisms of action and to the molecular basis of CS syndrome.</p></div

Western blotting demonstrating the significant increase of ATF4, CHOP and EIF2a-p after CSB knockdown.

<p>B) Graphs showing qRT-PCR analysis of CSB mRNA expression in SKNBE-2c cells, at 12h after the transfection. The results, normalized to β-actin, were averaged from values obtained by performing three technical replicates. The values are means ± SD. Graphs showing apoptosis (C) and cell viability (D) percentage in SKNBE-2c cells 48h after CSB ablation. The results were averaged from values obtained by performing three technical replicates. The values are means ± SD. E) Graphs show relative fold change values of up-regulation of selected genes in SKNBE-2c cells. The results, normalized to β-actin, were averaged from values obtained by performing three technical replicates. The values are means ± SD.</p

Venn diagrams showing pair-wise comparison between ASO and CTRL, ASO and OLF, finally ASO and SO at 6h and 12h.

<p>For intersections letters are in clockwise order of, for example DD for {Anti vs Ctrl} Intersect {Anti vs Oligo} means <i>first</i> D in {Anti vs Ctrl} and <i>second</i> D in {Anti vs Oligo}. Concerning the central intersection (yellow numbers) the clockwise order starts from Anti vs Ctrl. D means down-regulated and U up-regulated.</p

Additional file 2: Table S1. of RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells

Top RISC-released/loaded genes and top ribosome-loaded/released genes during the ES–ELA transition. (XLS 260 kb

Rats used for microarray analysis exhibited motor behaviors similar to the behaviors of the whole rat group.

<p><i>A,</i> The quality of the reaching behavior of the 4 rats in the <i>12-day-Reach</i> group (red, mean±SE) showed much improvement over time with the percentage of successful trials with dropped pellets of Day 8 to 12 being clearly lower than that of Day 1 to 5 (*, p<0.05, ANOVA). Similarly, a decreasing trend of this measure of learning was also observed in the <i>5-day-Reach</i> group (green). In this figure we have normalized the learning curve of every rat to its maximum value to account for inter-individual variability in initial performance. <i>B,</i> The probability of successful pellet retrieval per reach for the rats in the <i>12-day-Reach</i> group over the Learned (red, mean±SE) and Not-Learned (blue) Slots. Similar to the behavioral trend of the whole rat group (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061496#pone-0061496-g002" target="_blank">Fig. 2D</a>), the learning curve for the Learned Slots exhibited a sigmoid time course, with performance starting to increase at Day 5.1±3.2 (t_10%-max, black dotted line, mean±SE), and with the probability values of Day 1 to 5 smaller than those of Day 8 to 12 (*, p<0.05, ANOVA). The success probability values for the <i>5-day-Reach</i> group (N = 4; green; all slots) were also not statistically different from the values for the <i>12-day-Reach</i> group over the first 5 days (p>0.05, ANOVA).</p