(A) Primary mouse hepatocytes were infected with adenovirus-COX8-GFP-mCherry (multiplicity of infection [MOI] = 10) from seeding for overnight. Cells grown on coverslips were fixed in 4% paraformaldehyde (PFA) at indicated time points followed by LAMP1 immunofluorescence staining. Representative images from confocal microscopy analysis are shown. Arrows denote the colocalization of red-only mitochondria from COX8-GFP-mCherry assay with the lysosomal marker LAMP1.
(B) Quantification data for the number of red-only mitochondria in each cell from COX8-GFP-mCherry assay. Data are presented as mean ± SD. At least 15 fields from 2 or 3 independent mouse hepatocytes isolation were quantified.
(C) Schemes for type I and type II LAMP1-associated mitochondrial structures.
(D) Representative images of immunogold electron microscopy using an anti-LAMP1 antibody from day 3 cultured mouse hepatocytes.
(E) Total cell lysates from primary mouse hepatocytes cultured for indicated times were subjected to western blot analysis. *p < 0.05, two-tailed Student’s t test.