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. 2016 Aug 19;8(51):88386–88400. doi: 10.18632/oncotarget.11393

Figure 3. SH003 induces autolysosome formation.

Figure 3

(A) Stable expression of mCherry-GFP-LC3 MDA-MB-231 cells were treated with 500 μg/ml of SH003 for 24 hours and images were obtained with using Olympus FV10i Self Contained Confocal Laser System. Yellow (double staining with GFP and RFP) and red (staining with only RFP) florescence were stained for autophagosome and autolysosome, respectively. The object was 20× and scale bar indicates 10 μm. *P < 0.05. (B) MDA-MB-231 cells were treated with SH003 for 24 hours and stained with DND-99 lysotracker dye (75 nM) for 1 hour at 37°C. After fixation, permeabilization and blocking, cells were stained with anti-LC3B and Alexa-488 antibodies. DAPI was used for nucleus staining. (C) Cells were treated with SH003 for 24 hours and stained with anti-LC3B and LAMP1 (1:100) antibodies. (D) MDA-MB-231 cells were stained with anti-LC3B and LAMP2 (1:100) antibody. Colocalization with LC3B and LAMP2 was analyzed using Olympus FV10i Self Contained Confocal Laser System. The object was 20× and scale bar indicates 10 μm. *P < 0.05.