Figure 5. Cat S deficiency increases the Ang II-induced accumulation of autophagosomes in mouse heart.
Immunostaining for LC3 and F4/80 in (A) mouse heart (Bars, 10 µm) and (B) macrophages (Bars, 5 µm). (C) Western blot analysis of LC3 protein expression in macrophages. (D) RAW 264.7 cells were transfected with Cherry-GFP-LC3, and treated with and without Ang II in the presence or absence of Cat S inhibitor (15 nmol/L). Confocal fluorescence was used to image autophagosomes (red and green foci) and autolysosomes (red-only foci). Bars, 5 µm. (E) Representative electron microphotographs of cytoplasmic vacuolization in macrophages treated with saline or Ang II (100 nM) for 48 hr (n = 3 per group). Bars, 1 µm. (F) Immunostaining for LC3 and mitochondria marker MitoTtracker in macrophages. Bars, 2 µm. Data are mean±SEM (n = 4 per group). *P<0.05 vs. Cat S KO control macrophages. ## P<0.01 vs. Ang II-treated WT macrophages. §§ P<0.01 vs. WT control macrophages.