Maternal protein restriction induces renal AT2R promoter hypomethylation in salt‐sensitive, hypertensive rats

Figures

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  FIGURE 1. Negative correlation between AT2R DNA methylation and expression levels. (a) Schematic drawing of constructed reporter plasmid, indicating restriction enzyme sites of SmaI and XbaI. (b) Photo of agarose gel electrophoresis. Reporter plasmids with different DNA methylation levels were digested with SmaI and XbaI, then examined using agarose gel electrophoresis. Non‐Me, non‐methylated plasmid; L‐Me, low‐methylated plasmid; M‐Me, moderately methylated plasmid; and H‐Me, highly methylated plasmid. Marker is a HindIII‐digested λ DNA. (c) Photinus luciferase activity of reporter plasmids, normalized against Renilla luciferase activity as a control. Data represent means ± SEM (n = 3)

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  FIGURE 2. Effect of maternal and grand‐maternal protein restriction on blood pressure. (a) Blood pressure of 12‐week‐old offspring in the water drinking group. (b) Blood pressure of 12‐week‐old offspring in the salt loading group. Values are means ± SEM (n = 5–7). *p < .05 versus. control, one‐way ANOVA and Tukey's test. Offspring and grand‐offspring were exposed to either a control 20% casein diet or a 9% casein diet in utero. In adulthood, they were provided with either water (W) or 1% saline (S) to drink. CON, control; MPR, maternal protein restriction; GPR, grand‐maternal protein restriction; W, water; S, saline

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  FIGURE 3. Renal DNA methylation profiles in AT2R promoter region. (a) Schematic profiles of AT2R promoter region. Renal AT2R DNA methylation profiles of the water drinking (b) and salt loading (c) groups. Values are means ± SEM (n = 3). *p < .05, one‐way ANOVA and Tukey's test. Offspring were exposed either a 20% (CON) or 9% (MPR)‐casein diet in utero, and provided with either water (W) or 1% saline (S) to drink in adulthood

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  FIGURE 4. Effect of maternal and grand‐maternal protein restriction on AT2R protein level. Renal AT2R protein levels in the water drinking (a) and salt loading (b) groups. Values are means ± SEM (n = 5–7). Offspring and grand‐offspring were exposed either a 20% (CON) or 9% (MPR, GPR) casein diet in utero, and provided with either water (W) or 1% saline (S) to drink during adulthood

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  FIGURE 5. AT2R mRNA expression measured using digital PCR. (a) Water drinking groups. (b) Salt loading groups. *p < .05 versus. CON‐S, one‐way ANOVA and Tukey's test. Values are means ± SEM (n = 5–7). Offspring and grand‐offspring were exposed either a 20% (CON) or 9% (MPR, GPR) casein diets in utero, and provided with either water (W) or 1% saline (S) to drink in adulthood