Figure 4.

FAM3D deficiency inhibits AngII-induced eNOS uncoupling in ECs of aortas in mice

(A) Representative DAF-FM DA en face staining and quantification of NO levels in ECs from the thoracic aortas of WT and FAM3D^−/− mice after saline or AngII infusion for 14 days. n = 5–6. Data are represented as mean ± SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test, ∗p < 0.05.

(B) Representative DHE en face staining and quantification of O₂⁻ in ECs from thoracic aortas in WT and FAM3D^−/− mice infused with saline or AngII for 14 days. L-NAME (200 μmol/L) was ex vivo used to treat dissected aortas for 30 min before DHE staining. n = 3. Data are represented as mean ± SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test, ∗p < 0.05.

(C) Representative western blot analysis and quantification of monomer/dimer eNOS ratios in the aortas of WT and FAM3D^−/− mice treated with saline or AngII for 14 days. n = 5. Data are represented as mean ± SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test, ∗p < 0.05.

(D) Representative DAF-FM DA en face staining and quantification of NO in ECs from the thoracic aortas of WT and FAM3D^−/− mice infused with AngII and administered with or without 2,4- DAHP (10 mmol/L) in drinking water for 14 days. n = 5. Data are represented as mean ± SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test, ∗p < 0.05.

(E) SBP was measured by the tail-cuff method in WT and FAM3D^−/− mice infused with AngII and administered with or without DAHP (10 mmol/L) in drinking water for 14 days. n = 4–5. Data are represented as mean ± SEM. Repeated-measures analysis using a mixed-effects model followed by Tukey’s multiple comparisons test, ∗p < 0.05, WT + AngII vs. FAM3D^−/− + AngII; ns, no significance, WT + AngII + DAHP vs. FAM3D^−/− + AngII + DAHP.