MOESM1 of Cerium oxide and barium sulfate nanoparticle inhalation affects gene expression in alveolar epithelial cells type II

Additional file 1: Table S1. Fold regulation of regulated genes. List of fold regulation and standard deviation values of the regulated genes listed in Tables 2 and 3 of the manuscript. Values are shown for every dose group and time point

Detection of viral antigen in the brain of BDV-infected animals.

<p>After 21 dpi, the viral nucleoprotein BDV-N showed a disseminated and comparable distribution in the entire brain in all TNF-transgenic and wild-type animals. Score for the BDV-N immunoreactivity: 0: no detection of antigen; 1: single foci with BDV-N positive cells in some brain areas, <80 cells per HPF; 2: less than 150 positive cells per HPF, neuropil reaction, 3: more than 150 positive cells per HPF, distinct neuropil reaction p.i.: post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, median, bar: minimal/maximal value.</p

Transgenic TNF (TNFtg) mRNA values in different brain areas in TNF-transgenic and wild-type mice.

<p>TNFtg mRNA was present only in the transgenic animals. TNFtg mRNA copy numbers were significantly higher in the cerebral cortex, hippocampus and striatum when compared to the cerebellum in Tg/– and Tg/Tg animals regardless of BDV-infection. No TNFtg mRNA was detected in the wild-type mice in any brain area. dpi: days post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, geometric mean, bar: distribution factor</p

Frequency of seizures in BDV-infected mice.

<p>Spontaneous epileptic seizures were exclusively observed in BDV-infected transgenic animals and appeared more frequently in homozygous transgenic BDV-infected mice starting 21 dpi. Seizures were first noted 42 dpi in heterozygous transgenic animals. dpi: days post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice</p

Scoring of the clinical-neurological examination.

<p>Scoring of the clinical-neurological examination.</p

Development of weight gain in mock-infected and BDV-infected mice.

<p>TNF-transgenic animals gained less weight than the wild-type mice after BDV-infection. In general, all BDV-infected mice groups exhibited al lower weight gain when compared to mock-infected animals. dpi: days post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, BDV: BDV-infected, mock: mock-infected, arithmetic mean.</p

Degree of inflammatory reaction in the brain of BDV-infected animals.

<p>Wild-type mice showed a mild immune cell infiltration without progression during the investigation period whereas in both TNF-overexpressing transgenic mice groups a progressive severe non-purulent meningoencephalitis was noted. Highest inflammatory scores were found in the homozygous mice. Score for the inflammatory reaction: 0: no, 1: mild, 2: moderate, 3: severe inflammatory reaction; score for the astrogliosis: 0: no, 1: mild, 2: moderate, 3: severe astrogliosis, p.i.: post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, median, bar: minimal/maximal value.</p

Total TNF (TNFto) mRNA values in different brain areas in TNF-transgenic and wild-type mice.

<p>TNFto mRNA values were significantly higher in transgenic mice and consisted of approximately 50% native TNF mRNA indicating that TNFtg induced native TNF mRNA expression. Highest copy numbers were found in homozygous mice. No significant increase of any TNF mRNA was detected in transgenic animals after BDV-infection but values were highest in the hippocampus. dpi: days post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, geometric mean, bar: distribution factor.</p

NR2B mRNA values in different brain areas in TNF-transgenic and wild-type mice.

<p>Neither transgene expression nor BDV-infection caused significant changes in NR2B mRNA expression. dpi: days post infection, –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice, geometric mean, bar: distribution factor.</p

Inflammation, glial activation and viral distribution in the brain of BDV-infected TNF-transgenic and wild-type mice.

<p>A–F Hematoxylin and eosin staining: Mild inflammatory reaction and no microglia activation in non-transgenic BDV-infected mice (A, higher magnification in D). In contrast, moderate to severe inflammatory reaction and microglia activation in heterozygous (B, higher magnification in E) and homozygous (C, higher magnification in F) TNF-transgenic mice. TNF-overexpressing striatum, 42 dpi Hematoxylin and eosin staining was used to assess the degree of encephalitis and number of reactive microglia cells. <b>G–I GFAP-immunostaining:</b> Astrogliosis in all BDV-infected mice groups with more GFAP-positive astrocytes in the transgenic animals. GFAP-immunostaining was carried out to assess the number and morphology of activated astrocytes as GFAP expressing cells. TNF-overexpressing striatum, 42 dpi –/– <b>J–L Immunostaining for the viral nucleoprotein: </b>Comparable viral distribution within the brain in all BDV-infected mice groups. Immunostaining for the viral nucleoprotein was used to assess the number of BDV-infected cells. TNF-overexpressing striatum, 42 dpi –/–: non-transgenic mice, Tg/–: heterozygous transgenic mice, Tg/Tg: homozygous transgenic mice,scale bars: 100 µm (A–C; G–L); 50 µm (D–F).</p