The mechanism of internalization study of QDgreen−CD−FA−C−2028 conjugate at IC80 value to cancer (H460, Du-145, and LNCaP) and normal (MRC-5 and PNT1A) cells

The influence of different endocytosis inhibitors on the internalization of QDgreen−CD−FA−C−2028 conjugate at IC80 value in cancer (H460, Du-145, and LNCaP) and normal (MRC-5 and PNT1A) cells. First, the cells were preincubated with: drug-free medium (no inhibitor), at 4 °C, 5 µM Cytochalasin D, 30 µM Amiloride, 80 µM Dynasore, 25 µM Pitstop 2 and 1.5 µM Filipin III for 30 min, followed by further incubation with QDgreen−CD−FA−C−2028 for 4 h. The internalization of QDgreen−CD−FA−C−2028 in cells was explored by Confocal Laser Scanning Microscopy (63× magnification; ZEISS LSM T-PMT). Based on the fluorescence properties of these compounds, green and orange fluorescence are representative for QDgreen and C−2028, respectively. The imaging conditions were: QDgreen (excitation 300 nm, emission 543 nm), C−2028 (excitation 528 nm, emission 553 nm). The scale bar is 20 μm.