MOESM1 of Preliminary paleomagnetic and rock magnetic results from 17 to 22Â ka sediment of Jeju Island, Korea: Geomagnetic excursional behavior or rock magnetic anomalies?

Additional file 1. This includes a figure comparing paleomagnetic inclination data for East Asia from some previous literatures and this study (Figure S1), available AMS 14C age data of the Gosan Formation (Table S1), some rock magnetic properties data (Table S2), AF-derived paleomagnetic direction data (Table S3), TH-derived paleomagnetic direction data (Table S4), and the estimated age-depth model data (Table S5) for the GSDS site of the Gosan Formation

Morphology and internal structure of yeast cells after plasma treatment.

<p>(<b>A</b>) Cells analyzed by scanning electron microscopy. (<b>B</b>) Cells analyzed by transmission electron microscopy. Yeast cells in water, saline, and YPD were exposed to Ar gas and plasma for 3 min and used for the microscopic analysis.</p

Bisulfite sequencing of CpG sites in the upstream region of the <i>PTN</i> gene.

<p>[A] Schematic of the four CpG sites in the promoter region of the PTN gene are indicated by the heavy black vertical lines. The numbers on the line indicate positions relative to the transcription start site. Legend: a, complex contains E2F and p107 (E2F+p107); b, upstream stimulatory factor (USF also known as gamma-factor). [B] The CpG methylation status in the upstream region of the <i>PTN</i> gene was analyzed in normal and ovarian cancer cells from hens by bisulfate sequencing. Each circle indicates a CpG site in the primary sequence, and each line of circles represents analysis of a single cloned allele. Closed and open circles are methylated and unmethylated CpGs, respectively. [C] Comparison of the sequences around CpG regions of <i>PTN</i> genes of chicken, mouse, and human. The 5′ flanking region (about 2.2 kb) of mouse and human <i>PTN</i> was compared to that of chicken <i>PTN</i> to identify differences in sequences around each CpG sites among those species.</p

Viability of yeast cells measured by colony formation and MTT assay after plasma treatment.

<p>(<b>A</b>) Relative percentage of colony formations by yeast cells in water, saline, and YPD after treatment with plasma, compared to Ar gas only. Relative colony forming units (CFU) are calculated as follows: relative CFU (%) = (CFU number of plasma treated yeast/CFU number of Ar gas treated yeast)×100. All value are the average of three technical replicates. (<b>B</b>) The relative level of formazan product measured as absorbance at 490 nm in cells treated with plasma in water, saline, and YPD, compared to that in Ar gas treated cells. Yeast cells were treated with plasma and Ar gas for 3 min. Relative absorbance was calculated as relative absorbance (%) = (absorbance at 490 nm in plasma treated cells/absorbance at 490 nm in Ar gas treated cells)×100. Each value is the average of three technical replicates.</p

Expression of PTN in the chicken oviduct.

<p>[A] Results of RT-PCR analysis using cDNA templates from each segment of the chicken oviduct with chicken <i>PTN</i> and chicken <i>ACTB</i>-specific primers. [B] <i>In situ</i> hybridization analyses of <i>PTN</i> mRNAs in the chicken oviduct. Cross-sections of the infundibulum, magnum, isthmus and shell gland of the chicken oviduct were hybridized with antisense or sense chicken <i>PTN</i> cRNA probes. [C] Immunoreactive PTN protein in the chicken oviduct. For the IgG control, normal rabbit IgG was substituted for the primary antibody. Sections were not counterstained. Legend: LE, luminal epithelium; GE, glandular epithelium. <i>Scale bar</i> represents 100 µm. <i>See </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034215#s4" target="_blank"><i>Materials and Methods</i></a><i> for complete description</i>.</p

Phosphorylation of <i>HOG1</i> MAP kinase after Ar gas and plasma treatment analyzed by Western blot.

<p>The levels of Hog1p expression and phosphorylation are shown in the first and second panel, respectively. The third panel shows the Western blot for actin as loading control. Total protein was extracted from the cells treated with Ar gas and plasma in water, saline, and YPD for 3 min, and run on a 12% SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) gel.</p

Quantitation of expression of PTN and ESR1 in normal and cancerous ovaries from hens.

<p>[A] RT-PCR analyses were performed using cDNA templates from normal and cancerous ovaries of laying hens using chicken <i>PTN</i> and <i>ACTB</i>-specific primers. Lanes 1 to 4 show results of analyses of four normal ovaries. [B] Lanes 1–9 are from analyses of 9 different cancerous ovaries from laying hens. Expression of <i>PTN</i> mRNA was abundant in all carcinomas, but not in normal ovaries. Legend for panel B: Lane 1, endometrioid/serous/mucinous carcinoma (Stage III); Lane 2, endometrioid carcinoma (Stage I); Lane 3, serous carcinoma (Stage I); Lane 4, mucinous/endometrioid carcinoma (Stage IV); Lane 5, endometrioid carcinoma (Stage IV); Lane 6, endometrioid carcinoma (Stage III); Lane 7, clear cell carcinoma (Stage IV); Lane 8, serous/mucinous carcinoma (Stage IV); and Lane 9, serous/mucinous/endometrioid carcinoma (Stage III) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034215#pone.0034215-Lim1" target="_blank">[21]</a>. [C] The q-PCR analysis for <i>PTN</i> mRNA was performed using cDNA templates from normal and cancerous ovaries of laying hens (mean ± SEM; P<0.001). [D] <i>In situ</i> hybridization analyses of <i>PTN</i> mRNA in normal and cancerous ovaries of hens. Cross-sections of normal and cancerous ovaries of hens hybridized with antisense or sense chicken <i>PTN</i> cRNA probes demonstrated abundant <i>PTN</i> mRNA predominantly in GE of cancerous ovaries, but not in LE, stroma or blood vessels. [E] Immunoreactive PTN protein in normal and cancerous ovaries of hens. For the IgG control, normal rabbit IgG was substituted for the primary antibody. Sections were not counterstained. Legend: GE, glandular epithelium. <i>Scale bar</i> represents 200 µm (the first horizontal panels, sense and IgG) or 50 µm (the second horizontal panels, sense and IgG). [F] The q-PCR analysis for expression of <i>estrogen receptor alpha (ESR1)</i> was performed using cDNA templates from normal and cancerous ovaries of laying hens (mean ± SEM; P<0.001). [G] Immunoreactive ESR1 protein in normal and cancerous ovaries of hens. For the IgG control, normal rabbit IgG was substituted for the primary antibody. Sections were not counterstained. Legend: GE, glandular epithelium. <i>Scale bar</i> represents 200 µm (the first horizontal panels, sense and IgG) or 50 µm (the second horizontal panels, sense and IgG). <i>See </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034215#s4" target="_blank"><i>Materials and Methods</i></a><i> for a complete description of the methods</i>.</p

The level of reactive oxygen species (ROS) and reactive nitrogen species (RNS).

<p>ROS and RNS levels were measured on the surface of solutions (<b>A</b>), and inside solutions (<b>B</b>) and cells (<b>C</b>) after Ar gas and plasma treatments. Absorbance values in (A) were measured by optimal emission spectroscopy and are the average of five replicates. In (B) and (C), all values are normalized, in which the level measured after Ar gas treatment was subtracted from that measured after plasma treatment. Each value is the average of three technical replicates.</p

pH change after plasma treatment and effects of osmotic strength of background solutions.

<p>(<b>A</b>) pH of water, saline, PBS, and YPD after plasma treatment (left panel) and the effect of PBS on yeast cell viability (right panel). For pH measurements, 1 ml of each solution was exposed to plasma for the indicated time. The effect of PBS was assessed by measuring relative viability of plasma-treated cells compared to that of Ar gas-treated cells. Cells were treated with plasma for 3 min. (<b>B</b>) Viability of yeast cells in different concentrations of NaCl (left panel) and sorbitol (right panel) after plasma treatment. Yeast cells were treated with Ar gas and plasma for 1 and 3 min. (<b>C</b>) Changes in pH of NaCl (left panel) and sorbitol (right panel) solutions during plasma treatment. One ml of saline and sorbitol was exposed to plasma at the indicated concentrations for 10, 30, 60, and 180 sec. In (A) and (B), cell viability was assessed by relative ratio in colony forming units (CFU) calculated as follows: relative CFU (%) = (CFU number of plasma treated yeast/CFU number of Ar gas treated yeast)×100. All values are the average of three technical replicates.</p

Information on primers for RT-PCR analyses.

<p>Information on primers for RT-PCR analyses.</p