Size distribution of TiO₂ suspension in NaCl determined by dynamic light scattering.

<p>Size distribution of TiO₂ suspension in NaCl determined by dynamic light scattering.</p

Titanium dosage in organs and urine performed by ICP-OES.

<p>N.D. Not determined</p><p>Each value represents mean from 6 independent experiments ± SD. For each suspension, 3 measurements were performed.</p><p>(*) represent a statistical difference (p<0.05).</p><p>Titanium dosage in organs and urine performed by ICP-OES.</p

Transmission electron microscopy analysis and energy dispersive X ray microanalysis spectrum of TiO₂ agglomerates in urine, liver, spleen and kidneys 28 days after intravenous injection.

<p>Transmission electron microscopy analysis and energy dispersive X ray microanalysis spectrum of TiO₂ agglomerates in urine, liver, spleen and kidneys 28 days after intravenous injection.</p

HE optical microscopy analysisof TiO₂ agglomerates in target organs.

<p>(A) Liver, 1 day after treatment, (B) Liver, 56 days after treatment, (C) Lungs, 1 day after treatment, (D) Lungs, 56 days after treatment, (E) Spleen, 1 day after treatment, (F) Spleen, 56 days after treatment, (F) Kidney, 1 day after treatment, (G) Kidney, 56 days after treatment.</p

Data correlation in compartmental model.

<p>Data are represented as blue crosses. Means of data at each time are represented as pink dots. Confidence interval at 95% is represented by a dotted line (Mean ± 1.96 × SD). Results obtained with the compartmental model are represented as a black line.</p

Schematic representation of the kinetic models.

<p>Schematic representation of the kinetic models.</p

Histological analysis of 96 hpf larvae from DU-exposed adult fish compared to controls.

<p>(A-A’) Toluidine-blue staining of the larva at the level of the trunk. Vacuole-like structures (appearing as white dots) are abundant in DU-treated larvae compared to controls. (B-C) Mitochondrial morphology in skeletal muscles observed by transmission electronic microscopy of control larvae. The inner mitochondrial membranes are dense and well visible (black arrow head). (B’-C’) Alteration of mitochondrial morphology and absence or decreases density of inner mitochondrial membranes (*) in larvae from DU-exposed fish. (D-E) High-resolution transmission electronic microscopy of skeletal muscles in controls and (D’-E’) in the 96 hpf larvae obtained from DU-exposed fish. Disruption of myofibres (red asterisks) and swelling of Z-bands are visible in DU larvae. Z- and A-bands are indicated. Triads constituted of t-tubes and the two flanking cisterna of the sarcoplasmic reticulum are visible in the controls (black arrows) but are absent or deformed in larvae from DU-treated fish (red arrows).</p

Expression analysis of electron transport chain complex genes in testis and brain of DU exposed fish and in their progeny at two-cells stage and 96 hpf.

<p>(A) Heatmap of adjusted <i>p</i>-values of genes involved in mitochondrial oxydo-reduction process (<i>n</i> = 101). The colour code displays the log10(adjusted <i>p</i>-value) from the differential expression analysis. (B) MA-plot focusing on the differential expression of the electron transport chain complex genes in the 96 hpf larvae (fold change as DU/C). Most of the genes are up-regulated in DU exposed larva.</p

Differential expression analysis in two-cells stage embryos and 96 hpf larvae obtained from DU-exposed adult fish.

<p>(A) MA-plot at the two-cells stage. (B) MA-plot at 96 hpf. (C) Five-ways diagram of all genes regulated in adult brain, testis, ovaries and in the progeny of DU-exposed fish at two-cells stage and 96 hpf. (D) Examples of expression changes in the progeny of DU-exposed fish of genes involved in DNA repair (<i>rad51d</i>, <i>rad50</i>, <i>rad2</i>), splicing (<i>isy1</i>, <i>syf2</i>), chromatin remodelling (<i>hat1</i> and <i>hdac5</i>), lipid transport (<i>apoeb</i>, <i>cetp</i>), oxidative stress (<i>duox</i>) and cell adhesion (<i>pcdh1gc6</i>) (all adjusted <i>p</i>-values (FDR) < 0.01; error-bars represent the standard deviation to the mean).</p

Induction of protein chaperones expression following myofibre damage in the 96 hpf larvae from DU-exposed adult zebrafish.

<p>(A) MA-plot focusing on the differential expression of the 169 genes involved in cellular stress (from the ache mutant, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0177932#sec002" target="_blank">Material and methods</a>) and their regulation in 96 hpf larvae obtained from DU exposed. Significant adjusted <i>p</i>-values < 0.01 are indicated in red and yellow. (B) Log2 fold change of specific protein-chaperon in the skeletal muscles (all adjusted <i>p</i>-values (FDR) < 0.01; error-bars represent the standard deviation to the mean).</p