Using qPCR, we found that the selected genes expected to be upregulated by VPA in embryos [Lgals1, Mr2, and vinculin (Vcl)] or cells [keratin 1-18 (Krt1-18), Lgals1, and Mt2] were significantly (p < 0.05) induced (Figure 3).
Among these genes are several that were top ranked in embryos (Supplemental Material, Table 1), such as Mt2, metallothionein 1 (Mt1), Lgals1, H3f3b, creatine kinase-brain (Ckb), and transgelin 2 (Tagln2).
Among the six tested genes, the expression of Lgals1 and Vcl were significantly (p < 0.05) induced after 1.5 hr (Figure 5), which may support independently the VPA responsiveness of these two genes.
Along these lines, we found the expression of the gene Lgals1, which encodes the [beta]-galactoside-binding protein galectin-1 (Barondes et al.
Relative quantification (RQ) of genes ANXA1 and LGALS1 was performed according to the model proposed by Pfaffl , compared with pool of samples from normal gastric mucosa and normalized to the ACTB reference, according to the formula R = [(E target).sup.[DELTA]Ct target (control-sample)]/ [(E endogenous).sup.[DELTA]Ct endogenous (control-sample)].
However, for LGALS1, both groups showed basal gene expression, with overexpression in only 36.1% (13/36) of the IM cases (mean RQ = 0.35 [+ or -] 1.37) and 44.8% (13/29) of the GU cases (mean RQ = 0.69 [+ or -] 1.65) (Figure 1 and Table 2).
We also evaluated a possible association among the relative expression levels of ANXA1 and LGALS1 mRNA and the risk factors age, gender, smoking, drinking, H.
Regarding LGALS1, the relative gene expression presented basal values in both lesions.
The expression levels of the ANXA1, LGALS1, and ACTB mRNA were tested in triplicate (cDNA from the same RT reaction, but in separated wells).
Relative quantification (RQ) of genes ANXA1 and LGALS1 was obtained as described by Pfaffl  and normalized with the [beta]-actin control reference gene and normal gastric mucosa.
For LGALS1, the mRNA relative expression values found were lower; only the GA group showed overexpression in 60% (12/20) of the cases (mean RQ = 2.44 [+ or -] 3.26).