Figure 4

A transient increase of autophagy in rat liver. Changes in autophagic structures (day 1) (A), LC3-II aggregation (day 1–30) (B), and LC3-II protein expression (day 1–30) (C) in liver tissues at various time points following an i.p. Injection of Ag-nps. Experiments were carried out as described under Materials and Methods. A: TEM images of rat liver tissues. Scale bar = 0.5 μm. PAS = phagophore assembly site; Mit = mitochondria; AP = autophagosome; AL = autophagolysosome. B: upper panel: LC3-II aggregation was analyzed by immunofluorescence staining under fluorescence microscopy. Lower panel: quantitative analysis of FITC-LC3-II aggregation from each time points was estimated by counting the number of bright green dots, as arrows indicated (upper panel), under 200x amplification. Data were means ± SD of 8 individual rat experiment, *p < 0.05, **p < 0.01. C: LC3-II protein level changes based on Western blot analysis. Actin was used as a loading control. Values were expressed as means ± SD (n = 6), *p < 0.05 and **p < 0.01 versus sham group; ^# p < 0.05 versus Day 1 group.