Fig. 2: Elevation of NEP activity in Ensa KO mice.
A Immunostaining of NEP (Red) and VGAT (Green) from hippocampi of 3-month-old WT and Ensa KO mice. Scale bar is 100 µm in low magnification image and 50 µm in high-magnification image. B Statistical analysis of NEP immunoreactivity (n = 5 for each group). LM: lacunosum-molecular layer, Omo: Outer molecular layer and MMo: middle molecular layer. C Statistical analysis of colocalized NEP and VGAT signals (n = 5 for each group). D NEP activity in membrane fractions from hippocampi of 3-month-old WT and Ensa KO mice (WT: n = 7, Ensa KO: n = 6). E. Aβ40 ELISA of hippocampi of 3-month-old WT and Ensa KO mice (WT: n = 5, Ensa KO: n = 6). F. Aβ42 ELISA of hippocampi of 3-month-old WT and Ensa KO mice (WT: n = 5, Ensa KO: n = 6). G Aβ42 ELISA of hippocampi of 3-month-old Mme KO mice and Ensa/Mme dKO (Mme KO: n = 4, Ensa/Mme dKO: n = 5). H and I. Immunostaining of Aβ (Green), NEP (Red) and DAPI (blue) from 18-month-old AppNL-F and AppNL-F/Ensa KO mice. Statistical analysis of amyloid plaque area in 18-month-old AppNL-F and AppNL-F/Ensa KO mice (n = 6 for each group). Scale bar is 100 µm. J Aβ42 ELISA of Tris-HCl-buffered saline-soluble (Ts) hippocampal fractions from 18-month-old AppNL-F and AppNL-F/Ensa KO mice (AppNL-F: n = 7, AppNL-F/Ensa KO: n = 6). K Aβ42 ELISA of guanidine-HCl-soluble (GuHCl) hippocampal fractions from AppNL-F and AppNL-F/Ensa KO mice (AppNL-F: n = 7, AppNL-F/Ensa KO: n = 6). L Immunostaining of NEP (Red) and VGAT (Green) in hippocampi of 18-month-old AppNL-F and AppNL-F/Ensa KO mice. Scale bar is 100 µm in low-magnification image and 50 µm in high-magnification image. M Statistical analysis of NEP immunoreactivity (n = 4 for each group). N Statistical analysis of colocalized NEP and VGAT signals (n = 4 for each group). Data represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s or Welch’s t test).
