Autophagic Killing Effects against Mycobacterium tuberculosis by Alveolar Macrophages from Young and Aged Rhesus Macaques
Figure 1
Assessment of autophagy in alveolar macrophages from adult RM.
A. Western analysis was performed on cell lysates from resting control (C) or autophagic (Rap) alveolar macrophages using antibody against LC-3 and actin. The LC-3 antibody reacts with both free cytosolic LC3-I (18 kDa) and membrane associated LC3-II (16 kDa). Levels of LC3-II were normalized to actin by densitometry and the ratio of LC3-II/actin given below the blot. A representative image from a single rhesus macaque (RM) sample is shown. B. Immunofluorescence microscopy was performed on control and rapamycin-treated alveolar macrophages using primary antibody against LC-3. C. The number of cells possessing LC-3+ puncta (top) and the average number of LC3+ vacuoles in LC-3+ cells (bottom) were quantified (n = 6 NHP samples, n>50 macrophages in each condition). The difference between the number of cells possessing LC-3+ puncta was significantly different between control and autophagic cells (***, p<0.001; **, p<0.01; ANOVA).