Lysotracker green dnd

Doxorubicin hydrochloride (Dox·Hcl) was purchased from Beijing HuaFeng United Technology Co., Ltd. (Beijing, China). Resveratrol (Res) was bought from Guan Jie Biotech (Xian, China). 1-Bromo-4-chlorobutane and triphenylphosphine (TPP) were obtained from Aladdin Co., (Shanghai, China). K₂CO₃ and NaI were supplied by Sinopharm Chemical Reagent Co., (Shanghai, China). Dioleoylphosphatidylethanolamine (DOPE) and hydrogenated soybean phospholipids (HSPC) were purchased from Shanghai A.V.T Technology Co., Ltd. (Shanghai, China). Cholesterol and cholesteryl hemisuccinate (CHEMS) were bought from J&K Scientific Co., Ltd. (Beijing, China). 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), cell mitochondria isolation kit, mitochondrial membrane potential assay kit with JC-1, Caspase 3,8,9 assay kit, ATP assay kit, Lysotracker Green DND and Hoechst 33258 were purchased from Beyotime Biotechnology Co., Ltd. (Nantong, China). FITC anti-human CD44, FITC Mouse IgG1, PE anti-human CD24 and PE Mouse IgG2a were bought from BD Biosciences (Franklin Lakes, NJ, USA). β-Actin, β-catenin and cyclin D1 primary antibodies and corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies were provided by Abcam (Cambridge, UK).

KUP5 and AML12 cells were inoculated on confocal dishes and infected using the same process described in Sect. “Immunofluorescence of TFEB entry into KUP5 cells”. After CDs treatment, the supernatant was discarded and incubated with LysoTracker^® Red DND and LysoTracker^® Green DND at room temperature for 10 min and 15 min, respectively (Beyotime Technology, Shanghai, China). The red and green fluorescence signals were observed by confocal microscopy after fixation with 4% paraformaldehyde at 37 °C for 10 min.