Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for CRC in relation to genotypes of GRP78 polymorphisms.

a<p>Adjusted for age and gender.</p

Association of genotypes of GRP78 polymorphisms with clinicopathological characteristics of the CRC cases.

a<p>P was computed using chi-square test.</p><p>Among 414 CRC cases, 2 cases had missing data on TNM stage, 13 cases had missing data on tumor differentiation and 23 cases had missing data on tumor growth pattern.</p

Baseline demographic and clinical characteristics of CRC cases and controls.

a<p>P was computed using t test;</p>b<p>P was computed using chi-square test.</p><p>Among 414 CRC cases, 2 cases had missing data on TNM stage, 13 cases had missing data on tumor differentiation and 23 cases had missing data on tumor growth pattern.</p

Primers used to quantify mRNA.

<p>VEGF, vascular endothelial growth factor; ADRP, adipocyte differentiation-related protein; L-FABP, liver fatty acid binding protein; ALPI, intestinal alkaline phosphatase; F, forward primer; R, reverse primer.</p

PPAR δ knockdown promoted the growth of xenografts in nude mice.

<p>(A). Growth curve of xenografts. At each time point, the xenografts in PPAR δ-silenced group (n = 12) grew significantly larger than those of control group (n = 12) (*<i>P</i><0.05), even when treated by bevacizumab (**<i>P</i><0.05) (mean±SD, t-test). (B). Representative photograph of mice in each group was taken 25 days after inoculation. (C). The xenografts when nude mice sacrificed. (D) The xenografts in PPAR δ-silenced group were significantly heavier than those in control group when nude mice sacrificed (mean±SD; *<i>P = </i>0.021; **<i>P = </i>0.02; t-test).</p

The expression of Ki67 in xenografts.

<p>(A). Representative photographs of Ki67 expression in xenografts stained by IHC.×400 magnification. (B). PPAR δ- silenced group (n = 12) had significantly higher score of Ki67 staining than control group (n = 12) (*<i>P</i> = 0.026), even after treated by bevacizumab (**<i>P</i> = 0.031) (mean±SD; t test). (C). The mRNA level of Ki67 was significantly higher in PPAR δ-silenced group than in control group (*<i>P</i> = 0.018), even after treated by bevacizumab (**<i>P</i> = 0.025).</p

PPAR δ knockdown induced less differentiation of xenografts.

<p>(A). Representative photographs of xenografts by HE staining. The xenografts in PPAR δ-silenced group (n = 12) were obviously less-differentiated than those in control group (n = 12).×400 magnification. (B). PPAR δ- silenced group had significantly more less-differentiated xenografts than control group (85% vs. 17%, <i>P</i> = 0.025; Chi-square test). (C). Shown by quantitative RT-PCR, the mRNAs encoding ADRP, L-FABP or ALPI were significantly decreased in PPAR δ-silenced group relative to control group (<i>P</i><0.05).</p

KM12C cells secreted less VEGF after PPAR δ activation.

<p>The KM12C cells were treated with serial concentrations of GW501516 or vehicle, and the cell-free supernatants were collected for VEGF quantification by ELISA. Treated by GW501516, the control cells showed a dose-dependent decrease of VEGF secretion (*<i>P</i> = 0.018), while the PPAR δ-silenced cells had no significant change all along (**<i>P</i> = 0.83; ANOVA).</p

PPAR δ knockdown had no effect on apoptosis detected by TUNEL assay in xenografts.

<p>(A). Representative pictures of each group. The total cells were identified by DAPI stain, and the positive apoptosis cells were identified by FITC stain.×200 magnification. (B). Quantitative data of apoptosis index (AI). There wasn’t significant difference of AI between groups even after treated by bevacizumab (mean±SD,*<i>P</i> = 0.81, **<i>P</i> = 0.75; t-test).</p

PPAR δ knockdown increased VEGF expression in xenografts.

<p>(A). Representative photographs of VEGF expression in xenografts stained by IHC.×400 magnification; (B). PPAR δ- silenced group (n = 12) had significantly higher score of VEGF staining than control group (n = 12) (*<i>P</i> = 0.028; rank sum test). (C). Quantitative RT-PCR result (*<i>P = </i>0.032).</p