The <i>MTDH</i> (−470G>A) genotype & Allele distribution in Ovarian Cancer and controls.

†<p>The X² for HWE of Ovarian Cancer group and control group is 0.1 and 3.94 respectively (both <i>P</i>>0.05).</p

Association of the −470G>A genotype and <i>MTDH</i> (−470G>A) protein expression.

<p>A, Relative level of MTDH protein expression in ovarian cancer tissues compared to normal ovarian tissues. B, Relative level of MTDH protein expression in the ovarian cancer tissues of patients with different −470G>A genotypes. C, Relative level of MTDH protein expression in normal tissues of individuals with different −470G>A genotypes. One circle represents the mean of three independent measurements from one patient. The distribution of the three genotypes were random between the groups. N represents the samples number of respective group. Bars represent the standard deviation. Student’s t test was used to evaluate the differences in the expression levels of different constructs.</p

Sequencing chromatograms of <i>MTDH</i> (−470G>A).

<p>A–C, the sequencing chromatogram results of the genotype GG, GA and AA respectively. Samples were chosen randomly.</p

Knockdown of MTDH abolishes LPS-induced breast cancer cell invasion.

<p>(<b>A</b>) Images were taken from invasion assays using transwell system with or without the stimulation of LPS (100 ng/ml) for 18 hours in T47D, prp-control and prp-MTDH MDA-MB-231 cells. Images are shown at the original magnification of ×100. (<b>B</b>) The number of successfully invaded cells. The cell numbers were counted at five representative fields. Data was presented as mean±SD. The invaded cell number difference between T47D cells without LPS stimulation (22.6±2.1) and with LPS stimulation (25.2±1.9) was not significant (p = 0.074). The invaded cell number difference between prp-control MDA-MB-231 cells (92.2±8.6) and LPS-treated prp-control MDA-MB-231 cells (129.8±12.6) was significant; so was that between prp-control and prp-MTDH MDA-MB-231 cells with or without LPS stimulation, *:p<0.05 was considered significant compared with the LPS untreated cells.</p

MTDH regulates LPS-induced NF-κB activation and cytokine production.

<p>(<b>A</b>) Protein expression levels were determined by Western-blot assays. Protein extracts were prepared from the prp-control and prp-MTDH MDA-MB-231 cells treated without or with LPS (100 ng/ml) for 1 hour. (<b>B, C</b>) The mRNA levels of IL-8 (<b>B</b>) and MMP-9 (<b>C</b>) were measured by Real Time RT-PCR. Total RNA was extracted from cells in with or without of LPS (100 ng/ml) for 24 hours. β-actin was used as an endogenous control. Each experiment was carried out three times independently.</p

The <i>in vitro</i> scratch assays show increased migration ability in LPS-treated MDA-MB-231 cells.

<p>(<b>A</b>) Relative MTDH mRNA levels and protein levels in MDA-MB-231 cells transfected with prp-control or prp-MTDH vector respectively were measured by Real Time RT-PCR and western blot, respectively. (<b>B</b>) Images of un-pretreated or 100 ng/ml LPS-pretreated prp-control and prp-MTDH transfected MDA-MB-231 cells were taken at 0 and 24 hours. Images are shown at the original magnification of ×100. (<b>C</b>) Cell motility was quantified by measuring the distance between the migrating cell boundaries. Data were presented as mean±SD, n = 5. Significant difference on wound distance was observed between prp-control-MDA-MB-231 (prp-control) and prp-MTDH-MDA-MB-231 (prp-MTDH) cells, LPS-pretreated and un-pretreated prp-control cells, LPS-pretreated prp-control and prp-MTDH MDA-MB-231 cells. No significant differences was observed between LPS-pretreated and un-pretreated T47D and prp-MTDH-231 cells. T-test was carried out to examine the differences, *:p<0.05 was considered significant compared with the LPS untreated cells.</p

<i>GADD45A</i> genotypes and ovarian cancer prognosis.

<p>A, Genotype TT+TC of <i>GADD45A</i> had longer relapse-free survival (P = 0.018). B, Genotype TT+TC of <i>GADD45A</i> had longer overall survival (P = 0.0093).</p

Results of correlations among Th subsets and clinical characteristics.

<p>The pearson or spearman correlation test was used for correlation analysis. A. A positive correlation was found between Th1 and Th17 cells (r = 0.435, <i>P</i> = 0.0162). B, C. A negative correlation was shown between Th1 or Th2 cells with peripheral WBC (r = -0.39, <i>P</i> = 0.041; r = -0.407, <i>P</i> = 0.039, respectively) in ND AML patients. D, E. A negative correlation was found between Th17/Treg and leukemic blast (r = -0.439, p = 0.06), Th1/Th2 and BM leukemic blast (r = -0.691, p<0.0001) in BM of ND patients.</p

Sequencing chromatograms of <i>GADD45A (1506T>C)</i>.

<p>A-C, the reverse complement sequencing chromatogram results of the genotypes TT, TC and CC, respectively. The samples were randomly chosen.</p

The percentage of Treg (CD4⁺CD25⁺ Foxp3⁺), Th17 (CD4⁺IL-17⁺) cells in representative patients with ND, CR, relapsed-refractory AML patients or in controls.

<p>A. Lymphocytes were gated by flow cytometry. B. The percentage of CD4⁺ lymphocytes. C, D, E, F. The percentage of Treg (CD4⁺CD25⁺ Foxp3⁺) cells in AML patients and controls. H, I, J, K. The percentage of Th17 (CD4⁺IL-17⁺) cells in AML patients and controls.</p