Fig. 8.

Lysosome, autophagosome fusion in HMA treated cells. ARPE cells were cultured in normal medium (untreated), in a complete medium without aminoacids (to induce autophagy), in the presence of 50 nM bafilomycin (to inhibit the lysosomal proton pump) or 40 μM HMA. Immunofluorescence was performed using DAPI staining (blue), anti-LC3 (green) and anti-LAMP 2 (red) antibody. Insets in the ‘merge’ panel represent a zoomed region of the merged picture. In ‘untreated cells’ only lysosomes (red) show a punctuate staining. In the absence of amino acids (W/O aa) red and green stained (lysosomes and autophagosomes respectively) merged. These stains are not overlapped in bafilomycin-treated cells neither in HMA-treated cells. To note: given that HMA is fluorescent in UV conditions autophagosomes appear white in the merged pictures, lysosomes are red. The nucleus stained with DAPI is less fluorescent than the vesicles and appears artificially unlabelled. Scale bar: 25 μM.