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Gene expressions were analysed with Taqman Fast Advanced Master Mix (Thermo Fisher Scientific) by quantitative PCR using Step One Plus (Applied Biosystems, Foster City, CA, USA).
Differences in mRNA expressions were analysed using t-tests.
Thereafter, MHC class I and HER-2 expressions were analysed by flow cytometry.
β -Catenin and COX-2 expressions were analysed using the quantitative RT-PCR and Western blotting.
For each gene, differences between cell culture expressions were analysed by a two-tailed unpaired t-test.
The correlations of I-LVD and P-LVD with clinicopathologic parameters and VEGFs expressions were analysed by independent samples t test or Mann-Whitney U test.
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Where required, the significance of differences between relative gene expressions was analysed by two-way ANOVA.
Relative changes in gene expression were analysed via the 2ΔΔC(T) method.
Protein phosphorylation and expression were analysed by immunoblotting (images with black bands) (a) and quantified (b).
Protein phosphorylation and expression were analysed by immunoblotting (c,g) and quantified by densitometry (d,h).
The relative changes in gene expression were analysed by the 2−ΔΔCT method [57].
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