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. Author manuscript; available in PMC: 2021 Jul 19.
Published in final edited form as: Assay Drug Dev Technol. 2016 Sep 22;14(8):489–510. doi: 10.1089/adt.2016.727

Figure 3. Detection of Lysosomal membrane permeabilization with Galectin-3 reporter cell lines.

Figure 3

A. Left Panel. Schematic representation of the mAG-Gal3 reporter assay. Cells express Gal-3 in the cytosol and contain intact lysosomes (back vesicles). After compound treatment, the rupture of the lysosomal membrane allows the entry of the cytosolic expressed mAG-Gal3 protein, resulting in bright fluorescent dots (permeabilized lysosomes). Right panel. Confocal images of MCF7-mAG-Gal3 cells, after treatment for 8hs with the indicated compounds.

B. MCF7-mAG-Gal3 cells were treated with 30 μM Siramesine for 6 h and then stained with Lysotraker Red-DND-99 to mark the intact lysosomes. Right panel. Zoom-in of the indicated area showing the single channels images for mAG-Gal3 and Lysotracker-Red, and merge. The arrows point to damage lysosomes that showed strong Gal-3 stain and were negative lysotracker. The arrowheads point to lysosomes that haven’t totally release their content yet, showing lysotracker staining and either negative or deem stain for mAG-Gal3.

C. Left Panel. Schematic representation of the mCherry-Gal3 reporter assay. The principle is as in A. Right panel. Confocal images of MCF7-mCherry-Gal3 cells, after treatment for 6 h with the indicated compounds.