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. 2022 Sep 2;50(4):130. doi: 10.3892/ijmm.2022.5186

Figure 3.

Figure 3

DB does not affect the pH or the hydrolytic function of lysosomes. (A) Fluorescence images of MCF7 and MDA-MB-231 cells transfected with double fluorescent mRFP-GFP-LC3 lentivirus and treated with DB, CQ alone or in combination for 6 h. Scale bars, 10 µm. (B) Fluorescence microscopy with AO staining. MCF7 and MDA-MB-231 cells were treated with 20 µM DB or 60 µM CQ for 6 h and then stained with AO. The red fluorescence represents the acidic vesicles. Scale bars, 20 µm. (C) Flow cytometry for LysoTracker Red in MCF7 and MDA-MB-231 cells treated with DB (0, 5, 10 and 20 µM) for 6 h. (D) Representative western blots and corresponding protein quantification plots of CTSD and CTSB protein expression in MCF7 and MDA-MB-231 cells treated with 20 µM DB for 6 h. Cofilin was used as a loading control. DB, dicitrinone B; RFP, red fluorescent protein; GFP, green fluorescent protein; LC3, microtubule associated protein 1 light chain 3; AO, acridine orange; CQ, chloroquine; DB, dicitrinone B; CTSD, cathepsin D; CTSB, cathepsin B.