Cell cycle assay kit pi rnase staining

THP-1 cells were washed with PBS, centrifuged, and immersed in cell cycle staining reagent (Dojindo). After 30 min incubation at 4°C in the dark, cells were washed with PBS analyzed by flow cytometry (BD). For cell cycle studies, THP-1 cells were washed with PBS, centrifuged, and resuspended in a working solution (Cell Cycle Assay Kit-PI/RNase Staining; Dojindo). The cells were incubated at 4°C for 30 minutes in the dark and then analyzed using a FACSort flow cytometer (BD, San Jose, CA). The ModFit LT program version 2.0 (Verity Software, Topsham, ME) was used to determine the percentage of cells in each stage of the cell cycle. For the surface protein expression of CD16/32, CD206, and CD163, the cells were labeled with phycoerythrin-labeled anti-human CD16/32, CD206, or CD163 antibodies (Biolegend, CA, USA) in a phosphate solution and incubated at 4°C for 30 minutes in the dark, then incubated with a buffered saline (PBS; GIBCO, CA, USA) containing 2% bovine serum albumin (BSA; Sigma, CA, USA) in PBS for 30 minutes. Dead cells were excluded based on propidium iodide staining.

THPC was purchased from Aladdin Chemistry Co., Ltd (Shanghai, China). HAuCl₄ and Na₂PdCl₄ were purchased from InnoChem Science & Technology Co., Ltd (Beijing, China). SH-PEG-OMe (Mw: 5000 Da), SH-PEG-FA (Mw: 5000 Da) and SH-PEG-Cy5.5 (Mw: 5000 Da) were purchased from Shanghai Tuoyang Biotechnology Co., Ltd. SOD assay kit, Cell Counting Kit-8 (CCK-8), Cell Cycle Assay Kit - PI/RNase Staining and Annexin V, FITC Apoptosis Detection Kit were purchased from Dojindo Laboratories (Kumamoto, Japan). MCD was purchased from Alfa Aesar. Pyrogallol was purchased from Energy Chemical (Shanghai, China). ABDA was purchased from Sigma-Aldrich. APF and HPF probes were purchased from AAT Bioquest Inc. SOSG probe was purchased from Thermo Fisher Scientific Inc. DNA Damage Assay Kit by γ-H2AX Immuno fluorescence and One Step TUNEL Apoptosis Assay Kit were purchased from Beyotime Biotechnology Co., Ltd (Shanghai, China).

For cell cycle assay, cells were collected and stained with propidium iodide (PI) using Cell Cycle Assay Kit - PI/RNase Staining (Dojindo, Japan) following manufacture’s protocol. For cell apoptosis assay, cells were treated with cDDP (10 μg/ml) or blank control for 24 h. Later, cells were labeled with Annexin-V FITC and PI using Apoptosis Detection Kit (Dojindo, Japan). LSRFortessa (BD) was then used for flow cytometry analysis.