2 4 amidinophenyl 6 indolecarbamidine dihydrochloride dapi solution

EdU staining was performed using the EdU Cell Proliferation Kit (C0075S, Beyotime) according to the product specification. Briefly, 1 × 10⁵ treated A549 and HCC827 cells were cultured into a 6-well plate for 24 h, then A549 and HCC827 cells were incubated with 15 μM EdU solution at 37 ℃ for 2 h, and then fixed with paraformaldehyde and redyed with 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) solution (C1005, Beyotime, Shanghai, China) for 10 min at room temperature, and finally observed with fluorescence microscope (Olympus, Japan).

To investigate the subcellular distribution of PSNPs, RAW 264.7 cells (1 × 10⁵ cells/mL) or J774A.1 cells (1 × 10⁵ cells/mL) were incubated with 50 µg/mL fPSNPs or Neil-red PSNPs for 4 h, followed by co-incubation with fluorescent dyes, such as MitoBright LT Red (Dojindo Molecular Technologies, Japan), LysoTracker Red, ER-Tracker Red, and 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate (Dil) (Beyotime, China) for 20 min. After washing with PBS, the cells were then stained with 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) solution (Beyotime) at room temperature for 10 min and subjected to confocal microscopy to measure the fluorescence intensities. Moreover, the localization and mitochondria ultrastructure change of PS in RAW264.7 cells and J774A.1 cells were evaluated by TEM (JEOL 1011, Japan).