PANC-1 cells treated with co-delivery NP at various loading ratios were also tested by BrdU proliferation assay. A series of PAL concentrations (0–20 μM) and HCQ concentrations (0–400 μM) were tested. Cells treated with HCQ NP and PAL NP at equivalent concentration were also tested. Cell proliferation was determined by BrdU proliferation assay kit (Cell Signaling Technology) as per the manufacturer’s instructions. The absorbance at 450 nm were normalized to saline control to give the percentage of proliferating cells. The results were analyzed with the CompuSyn software to obtain CI and IC50. To probe cell cycle, PANC-1 cells were treated with co-delivery NP (1:4.3 loading molar ratio, PAL = 2.5 μM, HCQ = 10.8 μM) for 72 h (n = 3). Cells were fixed with 4% paraformaldehyde for 10 min at room temperature. Then the cells were washed with phosphate-buffered saline (PBS) and incubated with Tali cell cycle solution (Thermo Fisher) that contains propidium iodide, RNase A, and Triton X-100 for 30 min in dark. To obtain single-cell suspension, the cells were passed through a 70-μm cell strainer. Cells were detected on a flowcytometer (LSRII, Becton Dickinson) with the BDFACSDivaTM software 7.0. The cell cycle analysis was performed with the FlowJo software (v7.6.1).
Tali cell cycle solution
Manufactured by Thermo Fisher Scientific
Sourced in China, United Kingdom, United States
The Tali cell cycle solution is a laboratory equipment product designed for cell cycle analysis. It provides a standardized and efficient method to determine the distribution of cells in different phases of the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Tali image based cytometer, by Thermo Fisher Scientific (2 mentions)
Facsdiva, by BD (1 mentions)
Brdu proliferation assay kit, by Cell Signaling Technology (1 mentions)
Annexin 5 pe 7 aad apoptosis detection kit, by Vazyme (1 mentions)
Fcs express research edition software, by De Novo Software (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Bio-Rad (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Merck Group (1 mentions)
4 protocols using tali cell cycle solution
1
BrdU Proliferation and Cell Cycle Analysis of Co-delivery Nanoparticles
2
Cell Cycle and Apoptosis Analysis of HCC
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After 48 h of HCC transfection, the cells were trypsinized and resuspended in DPBS. 75% precooled ethanol fixed the cells and washed it 3 times by DPBS before using for cell cycle analysis with Tali® Cell Cycle Solution (Thermo Fisher Scientific, Shanghai, China). The apoptosis of HCC cells were stained by Annexin V-PE/7-AAD Apoptosis Detection Kit (Vazyme Biotech Co., Ltd., Nanjing, China).
3
Cell Cycle Analysis of NRK-52E Cells
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Following 24 or 48 h high glucose treatment, NRK-52E cells were fixed in 70% ethanol and kept at −20°C. On the day of analysis, cells were pelleted out by centrifugation at 1000 ×g for 5 minutes at 4°C and stained with Tali® cell cycle solution (Life Technologies, UK) in dark for 30 minutes to achieve a cell density of 1 × 105–5 × 106 cells/ml for analysis using Tali image-based cytometer (Life Technologies, UK).
4
Cell Cycle Analysis of C2C12 Myoblasts
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Cell cycle analysis was performed as in Pierucci et al. [45 (link)]. Briefly, C2C12 myoblasts were collected, washed once with DPBS (Sigma Aldrich, Milan, Italy) and fixed with ice-cold 70% ethanol in dH2O (Sigma Aldrich, Milan, Italy) to a concentration of 1 × 106 to −5 × 106 cells/mL. The samples were placed at least for a night at –20 °C, then ethanol was removed, and cells were washed with DPBS + 1%BSA (Bio-Rad, Hercules, CA, USA). After DPBS was removed, cells were stained with propidium iodide incubating them at room temperature for 30 min in the dark with the Tali®® Cell Cycle Solution (Life Technologies, Eugene, OR, USA) to a concentration of 1 × 105 to –5 × 106 cells/mL. Cell suspension (25 µL) was transferred in the dedicated slide and cell cycle analysis was performed using the Tali®® Image-Based Cytometer (Thermo Fisher Scientific, Carlsbad, CA, USA). The percentage of cells in each phase of the cell cycle was determined using FCS Express Research Edition software (version 4.03; De Novo Software, Morristown, NJ, USA).
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