raw data of the study-Retrospective Study using Magnetic Resonance Imaging (MRI) to Measure Depths of Acupuncture Points in Neck and Shoulder Region

This document contains the raw data of 394 subjects including sex,BMI and measured depth of each point in the study: Retrospective Study using Magnetic Resonance Imaging (MRI) to Measure Depths of Acupuncture Points in Neck and Shoulder Region

Protection of K562 cells from lapatinib-induced cytotoxicity by knockdown of autophagy-related proteins.

<p>After transduction with shRNA expression lentivirus as indicated in each figure, K562 cells were selected and kept in puromycin-containing medium. Cells were treated with DMSO or lapatinib for 72 (A) or 48 h (B), and then the relative percentages of growth inhibition were detected using the MTS assay (A, left panel and B, upper panel) or trypan blue exclusion assay (A, right panel and B, lower panel) and calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029014#pone-0029014-g001" target="_blank">Fig. 1</a>. Knockdown efficiency of ATG7, beclin-1, and ATG5-12 conjugates or the loading control, actin, were examined by immunoblotting using antibody against the respective proteins as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029014#pone-0029014-g003" target="_blank">Fig. 3</a>. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 (<i>t</i>-test).</p

Induction of apoptosis by lapatinib in K562 and HL-60 cells.

<p>(A) K562 or HL-60 cells were left untreated or treated with various concentrations of lapatinib or TPA as indicated for 1–3 days. Cells were collected and resuspended in propidium iodide (PI)-containing hypotonic buffer, and then the percentage of apoptotic cells with DNA ladders (hypodiploid cells) was analyzed by flow cytometry. The data are expressed as means ± SEMs. (B) K562 cells were left untreated or treated with lapatinib for 3 days; cells were then collected, resuspended in both AnnexinV and PI containing buffer, and analyzed by flow cytometry. The percentages of PI(+)/annexin(+) or PI(−)/annexin(+) cells are indicated in each figure. (C) Induction of both apoptotic and non-apoptotic cell death by lapatinib in K562 cells. After DMSO or 10 µM lapatinib treatment for 1–3 days for K562, or 3 days for HL-60, cells were split into two tubes and resuspended in PI-containing phosphate buffered saline (upper panel in left figure, total dead cells) or PI-containing hypotonic buffer (lower panel in left figure, apoptotic cells), respectively, for simultaneously detecting total dead cells without intact plasma membranes or apoptotic cells as described in (A). The graph in the right panel represents the data as: the percentage of total dead cells (empty bars) and the percentages of apoptotic cells (solid bars). After drug treatment for 3 days, HL-60 cells were attached on slides using cytospin apparatus and observed after staining with Liu's stain (right panel of 2C). (D) After DMSO, 2.5, 5 or 10 µM lapatinib treatment for 8 or 16 h, K562 cells were stained with both PI and DiOC6₍₃₎. The mitochondrial transmembrane potential of the cells was analyzed by flow cytometry. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 (<i>t</i>-test) between treated and DMSO control cells.</p

Rescue of K562 cells from lapatinib-induced cytotoxicity by the autophagy inhibitor 3-methyladenine (3-MA).

<p>K562 cells were left untreated or were treated with various concentrations of lapatinib in the presence or absence of 1.25-mM or 2.5-mM 3-MA for 24 (A) or 48 h (B) as indicated in each figure. Relative amounts of viable cells were detected using the MTS assay, and the relative percentage of growth inhibition was calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029014#pone-0029014-g001" target="_blank">Fig. 1</a>. *<i>P</i><0.05, **<i>P</i><0.01 (<i>t</i>-test).</p

Inhibition of leukemia cell proliferation by lapatinib.

<p>K562 CML cells were left untreated or were treated with 0.1% dimethyl sulfoxide (DMSO, vehicle), DMSO with different doses of lapatinib (2.5, 5, or 10 µM), or 12-<i>O</i>-Tetradecanoylphorbol 13-acetate (TPA) 10 µM for 1 to 3 days as indicated. (A) Cell numbers for each treatment were counted using trypan-blue dye exclusion assay. (B) Relative numbers of viable cells were detected using MTT assay. (C–F) Inhibition of cell proliferation by lapatinib in leukemia cell lines, but not in primary CD14⁺ mononuclear or bone marrow cells. 1×10⁵/ml CML MEG-01 (C), AML NB4 (D), HL-60 cells (E), K562, or 5×10⁵/ml human CD14⁺ mononuclear and mouse bone marrow cells (F) were untreated (unTx), treated with DMSO vehicle (0), or DMSO with different doses of lapatinib (µM) for 2 (C), 3 (D–E), or 1–3 days (F). The raw data (B and F) or relative percentage of growth inhibition (C–E) were assessed and calculated using both trypan-blue dye exclusion and MTS assays (C–E) or MTT assays (F) as indicated in each figure as described in (A–B). Optical density (OD) values from DMSO-treated control cells were used as a standard (0% cell death), and the relative percentage of growth inhibition was calculated using the following method: [(mean OD values from DMSO-treated cells – mean OD values from drug-treated cells)/mean OD values from DMSO-treated cells]×100. The data are expressed as the mean ± SEM. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 (<i>t</i>-test) between lapatinib-treated and DMSO control cells.</p

Frequency of GNMT polymorphisms and association with prostate cancer risk.

<p>aOR = age-adjusted OR.</p><p>*Minor allele frequency in controls = 0.456.</p><p>**not in HWE.</p

Analysis of polymorphisms according to prostate cancer aggressiveness and lethality.

<p>aOR = age-adjusted OR, number of controls = 656.</p><p>*MAF in controls = 0.46.</p><p>**not in HWE.</p><p>p-heterogeneity between aggressive and non-aggressive for rs10948059: CT vs CC = 0.06, TT vs CC = 0.09, per-allele = 0.09.</p

Diagnostic sensitivity and false-positive identify for cancers provided by a single nucleoside and the set of six nucleosides.

<p>(A) All cancer; (B) Colon cancer, (C) Breast cancer; (D) Lung cancer and (E) Gastric cancer.</p

Clinical characteristic of colon and lung cancer patients.

<p>Clinical characteristic of colon and lung cancer patients.</p

Characteristics of prostate cases (PCa) and controls.

<p>* 20 missing data on stage (3.0%).</p><p>** 31 missing data on Gleason score (4.7%).</p><p>*** 44 missing data on PSA at diagnosis (6.7%).</p