Table 1.
Characteristics of the target gene sequences and primers condition.
Table 2.
Clinical characteristics of fifty pregnant women and their newborns from Mexico City.
Fig 1.
DNA methylation status in a single locus of GNPDA2 and PGC1α promoters from newborns.
(A) and (C) are representative difference plots of normalized and temperature-shifted gene promoters showing the reference curves of methylation as 0%, 1%, 10%, 50%, and 100% methylated universal standard dilutions. (B) and (D) are the gene promoter methylation statuses represented by melting peaks plots.
Fig 2.
GWG impacts on DNA methylation variability in a single locus of the LEPR promoter.
(A) Normalized and temperature-shifted difference plot. (B) Melting peaks plot of LEPR promoter methylation status, comparing it with the reference curve from human control sets as 0%, 1%, 10%, 50%, and 100% methylated universal standard dilutions. (C) LEPR methylation status expressed by the mean of AUC (DNA samples of 10% = 194.4 ± 40.9; of 1% = 259.3 ± 15.3; of 0% = 308.6 ± 17.8) and its comparison with the control sets (100% = 15.3 ± 5.3; 10% = 160.3 ± 64.3; 1% = 283.3 ± 69.7; 0% = 325.1 ± 59.9). Different letters over each box and whiskers show statistical difference data (p < 0.05) obtained by one-way ANOVA post-Tukey’s multiple comparisons test. (D) Negative correlation between LEPR methylation status and GWG obtained by Pearson’s correlation test. (E) Differential LEPR methylation status in the newborns that were classified by maternal total weight gain according to recommendations of the IOM. Statistical difference data (*p < 0.01) was obtained by Kruskal-Wallis test corrected by Dunn’s multiple comparisons test.
Fig 3.
LEPR methylation status variability in umbilical vein of newborns exposed to healthy and unhealthy maternal consumption habits.
Pie charts show the percentage of pregnant women who smoked, consumed alcohol, used drugs, and denied substance use, according to LEPR promoter methylation status.
Fig 4.
Site-specific LEPR promoter methylation status by Sanger sequencing.
(A) Methylation status profiles in WT, MS, human control sets (0, 1 and 10% of DNA methylation) and bisulfite-modified DNA samples by lollipop diagram representation. Methylated CpGs are represented in black lollipops, non-methylated CpGs are in white lollipops, and not defined CpGs are marked with X and asterisk. (B) Percentages of methylation obtained in each allele (forward and reverse sequence) from bisulfite-modified DNA samples and those compared to MS and control set percentages in both MS-HRM and Sanger methods.
Fig 5.
Lep-Rb expression in the umbilical vein from different statuses of gene promoter methylation at term pregnancy.
The figure shows a representative Western Blot and the signal quantification of Lep-Rb at three different DNA methylation status (0%, 1%, and 10%). The bars and error bars show the means ± SD of eleven protein extracts samples; difference between means (p < 0.05) was obtained by one-way ANOVA post hoc Tukey’s test and this is shown with different letters over the bars.