In vitro cytotoxicity assay was determined by CCK‐8 assay and PI staining assay. HCC cells were seeded into 96‐well plates at a density of 3000 cell/well and cultured overnight before treatment. Then, sorafenib, Gal‐LP and Gal‐SLP were added into medium at virous sorafenib and shUSP22 concentrations. After incubation for 48 h, cell viability was assessed by CCK‐8 assay as mentioned above. For PI staining, 100 000 HCC cells (Huh‐7 and BEL‐7402) were plated into 6‐well plates. Sorafenib, Gal‐LP and Gal‐SLP were added at a sorafenib concentration of 5 µм and shUSP22 concentration of 5 µg well−1. After treatment for 36 h, cells were harvested and stained with 5 µL PI solution (1 µg mL−1, KeyGen) for 15 min. PI‐positive cells proportion was quantitatively investigated by flow cytometer (BD FACSCanto II).
Pi solution
Manufactured by Keygen Biotech
Sourced in China
The PI solution is a laboratory reagent used in various analytical and experimental procedures. It serves as a buffer solution, maintaining a specific pH range to facilitate various biological and chemical processes. The core function of the PI solution is to provide a stable and controlled environment for samples or reactions, without making claims about its intended use or applications.
Automatically generated - may contain errors
Lab products found in correlation
Annexin 5 fitc, by Keygen Biotech (2 mentions)
Facscanto 2, by BD (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Annexin 5 apc 7 aad double staining apoptosis detection kit, by Liankebio (1 mentions)
Epics xl mcl, by Beckman Coulter (1 mentions)
Multimode plate reader, by Tecan (1 mentions)
Cellquest software, by BD (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
6 protocols using pi solution
1
Cytotoxicity Evaluation of Sorafenib and Nanoformulations
2
Cell Cycle Analysis of PVE Treatment
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TPC-1 and SW579 cells were incubated for 48 h in a complete culture medium containing PVE at 4, 2, or 0 mg/mL. Then, the cells were harvested, washed with cold PBS, and fixed in 70% v/v ethanol. After incubation for 6–8 h at 4°C, the cells were washed with PBS and suspended in 500 μl of a propidine iodide (PI) solution (Keygen, China) for 30 min before flow cytometry analysis. The percentages of cells in G1, S, and G2 phases were measured by flow cytometry (Beckman, CA, USA). The data were analysed using Multicycle-DNA cell cycle analysis software.
3
Cell Cycle Analysis by Flow Cytometry
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Cells (5×105 cells/well) with different treatment were seeded in six-well plates and serum starved for 24 h. Then washed with ice-cold phosphate-buffered saline (PBS), centrifuged, and fixed in ice-cold 75% (v/v) ethanol for overnight at 4°C. Then cells were suspended in propidium iodide (PI) solution (50 µg/mL) (KeyGEN BioTECH, Nanjing, China) with ribonuclease A (RNase A) (0.1 mg/mL) for 30 min in the dark. Cell cycle distribution was assessed using flow cytometer (Becton-Dickinson, San Jose, CA, USA).
4
Radiation-Induced Apoptosis and Cell Cycle Assay
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Cells were first seeded into a 6-well plate at a density of 1 × 105 cells per well and treated with 6-MV X-ray radiation at doses of 0 Gy and 8 Gy. Cells were collected and stained with an Annexin V-APC/7-AAD double staining apoptosis detection kit (LiankeBio, Hangzhou, China) according to the manufacturer's instructions. For cell cycle assays, cells were harvested and stained with PI solution (KeyGen Biotech, Nanjing, China) with RNase A. Flow cytometry was used to evaluate the luciferase intensity at 24 h after treatment.
5
Apoptosis Analysis in PASMCs
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At 48 h post-transfection, the PASMCs (1×106) were harvested, and 5 µl Annexin V-FITC (Key GEN Biotech Co., Ltd.) and 5 µl PI solution (KeyGEN Biotech Co., Ltd.) were added and stained for 15 min in the dark. The relative percentage of apoptosis was analyzed by flow cytometry (EPICS XL-MCL; Beckman Coulter, Inc.).
6
Cytotoxicity and Apoptosis of ATO on PBMCs
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Cytotoxicity of ATO on PBMCs was first evaluated using CCK-8 methods. Briefly, 196 μl of this PBMCs suspension was loaded into a 96-well round-bottomed plate at the density of 106 cells/ml. Then cells were treated with 4 μl ATO (1, 2.5, and 5 μmol/L) for 96 hours and incubated with 20 μl CCK-8 reagents for another 3 hours at 37°C. The optical density (OD) was detected at a wavelength of 450 nm on Multimode Plate Readers (Tecan, Mannedorf, Switzerland). The OD values for the experimental groups are represented as the percentage change compared with control.
For the cell apoptosis analysis, cells were harvested and washed with ice-cold PBS, and resuspended in 500 μl of binding buffer. Then cells were stained with annexin V-FITC (3.5 μl) and a PI solution (2.5 μl) for 15 minutes in the dark at room temperature (KeyGen, Nanjing, China). Flow cytometry analysis was performed using a FACScan flow cytometer (BD Biosciences, USA) and CellQuest software (Becton Dickinson, USA).
For the cell apoptosis analysis, cells were harvested and washed with ice-cold PBS, and resuspended in 500 μl of binding buffer. Then cells were stained with annexin V-FITC (3.5 μl) and a PI solution (2.5 μl) for 15 minutes in the dark at room temperature (KeyGen, Nanjing, China). Flow cytometry analysis was performed using a FACScan flow cytometer (BD Biosciences, USA) and CellQuest software (Becton Dickinson, USA).
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