Figure 4

Autophagy induced after stimulation with the NOD1 ligand in MDMs was productive. MDMs were transduced with p62-RFP during 18 h. Cells were incubated in the presence of 5 μg/ml of Tri-DAP for 24 h. Accumulation of p62 in autophagosomes (puncta) was evaluated by fluorescence microscopy (A). Lysotracker and Hoescht were used to stain lysosomes and nuclei, respectively. Choloroquine 60 μM was included as autophagic flux blocker. Transduction levels of p62 (5 h) and autophagy induction (24 h) were evaluated by counting cells with p62 puncta (B) and colocalization with lysotracker (C). The mean ± SE of two independent experiments is depicted. Up to 300 cells were counted for each condition.