Figure 1:

DC661 possesses exquisite anti-lysosome activity. (A) Chemical structures of HCQ, Lys05 and DC661. (B) A375P cells were treated with doses shown of HCQ, Lys05, or DC661 for 6 hours before lysates were immunoblotted. (C) A375P cells expressing the mCherry-eGFP-LC3B reporter were treated with HCQ, Lys05, or DC661 (0 – 100 μM, 1 hr) and analyzed by microscopy. (D) A375P cells were treated with HCQ, Lys05, or DC661 (3 μM, 6 hr) and stained with Lysosensor and imaged with fluorescent microscopy. Shown to the right is quantitation. (E) A375P cells were treated with HCQ, Lys05, or DC661 (3 μM, 6 hr) and subsequently stained with galectin-3 and imaged via fluorescent microscopy. Shown to the right is quantitation of Galectin-3+ puncta expressing cells. (F) A375P cells were treated chronically with HCQ, Lys05 or DC661 (0 – 1000 nM, 2 weeks) in a colony formation assay. Cells were subsequently stained with crystal violet and imaged. (G) HT-29 (colorectal cancer) cells were injected into the flanks of NSG mice. Once palpable, mice were treated daily with 3 mg/kg of DC661 or vehicle (water). Tumor volumes are shown. (H) Mean +/− SEM tumor growth rate. All data is representative from at least 2 experiments. * indicates p<0.05. Students T test was used.