Cell Cycle Analysis of Osteosarcoma Cells

All OS cell lines were analysed for cell cycle progression using Bromodeoxyuridine (BrdU) and Propidium iodide (PI) staining as previously described [34 (link)]. Briefly, OS cells were incubated for 72 h with medium containing vehicle or GIC₅₀ CX-5461 concentrations (determined from the cell proliferation assay), then labelled with 10 µM BrdU (Sigma-Aldrich, St. Louis, MO, USA), harvested and stained with PI. Flow cytometry (Becton Dickinson LSR II) was used to assess the level of staining, and FlowJo software (version 10.0) was used for data analysis.

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Kang C.W., Blackburn A.C., Loh A.H., Hong K.C., Goh J.Y., Hein N., Drygin D., Parish C.R., Hannan R.D., Hannan K.M, & Coupland L.A. (2023). Targeting RNA Polymerase I Transcription Activity in Osteosarcoma: Pre-Clinical Molecular and Animal Treatment Studies. Biomedicines, 11(4), 1133.

Publication 2023

BrdU LSR II

Assay Bromodeoxyuridine Cell cycle Cell lines Cell proliferation Cells Cx 5461 Flow cytometry Progression Propidium iodide

Corresponding Organization : Australian National University

Other organizations : KK Women's and Children's Hospital, Regulus Therapeutics (United States), University of Melbourne, Monash University, University of Queensland

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Variable analysis

independent variables

Treatment with vehicle or GIC50 CX-5461 concentrations

dependent variables

Cell cycle progression

control variables

Incubation time (72 h)
BrdU concentration (10 µM)
Propidium iodide staining

controls

Positive control: Not explicitly mentioned
Negative control: Vehicle treatment

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