Fig 2: The KD attenuated angiotensin converting enzyme 1 (ACE1) and angiotensin II, type I receptor (AT1R) contents in pulmonary tissue (A,C, respectively), however only the former was found to be statistically significant. Angiotensin II, type 2 receptor (AT2R) was unaltered in the lungs, regardless of diet (E). On the other hand, dietary intervention did not influence heart ACE1 content (B), however AT1R and AT2R were significantly elevated in the cardiac tissues of HFS-fed rats (D,F, respectively). Significant differences are denoted by different letters p < 0.05. Bars represent mean ± SEM. One-way ANOVA, n = 5–6.

Fig 3: The relationship between the lung expression and serum concentrations of ACE and ACE2(A) Correlation between serum ACE concentration and lung ACE mRNA level. (B) The absence of a correlation between serum ACE concentration and lung ACE protein abundance. (C) The absence of a correlation between lung ACE protein abundance and lung ACE mRNA level. (D) The absence of a correlation between serum ACE2 concentration and lung ACE2 mRNA level. (E) The absence of a correlation between serum ACE2 abundance and lung ACE2 protein abundance. (F) Correlation between lung ACE2 protein abundance and lung ACE2 mRNA level. The correlation coefficients (r) and P-values were obtained by Pearson’s correlation test, and lines were obtained by linear regression plotting.

Fig 4: Lung ACE and ACE2 mRNA and protein expression in male and female normotensive (Wistar) and hypertensive (SHR) ratsThe gene expression of (A) ACE and (B) ACE2 was quantitatively measured by RT-PCR and normalized to cyclophilin and is expressed as the % of male Wistar rats. (C) Lung ACE/ACE2 gene expression ratio was expressed as 2-?Ct/2-?Ct. The protein expression of (D) ACE and (E) ACE2 was determined by immunoblotting of lung membrane proteins and normalized to ß-actin and is expressed as the % of male Wistar rats (Supplementary Figures S3 and S4). The values represent individual measurements and the means ± SEMs. *P<0.05; **P<0.01; ***P<0.001.

Fig 5: Representative immunoblots (A) and protein expression analysis of ACE (B), Tapbp (C), and ß2m (D) in hippocampus. Representative immunoblots (E) and protein expression analysis of ACE (F), Tapbp (G), and ß2m (H) in olfactory bulbs. Data were analyzed by two-way ANOVA with Bonferroni post hoc test and presented as mean ± SEM (n = 3–6): * p = 0.05, ** p = 0.01.