Assaying Cellular Oxidative Stress and Lipid Peroxidation

OS cells, with or without applied genetic modifications, were placed onto 12-well plates at 0.5 × 10⁵ cells per well and cultivated for indicated time periods. Cells were then washed once with PBS and stained with the applied fluorescence dyes (DCF-DA or CellROX, dissolved in the medium). After extensive washes, fluorescence images were visualized under a fluorescence microscopy (Leica), and its intensity was detected by a fluorescence spectrophotometer (F-7000, Hitachi-Hightech). A thiobarbituric acid reactive substances (TBAR) activity assay kit was employed to examine cellular lipid peroxidation levels using the protocol described [21 (link), 22 (link)].

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Zhuo B.B., Zhu L.Q., Yao C., Wang X.H., Li S.X., Wang R., Li Y, & Ling Z.Y. (2022). ADCK1 is a potential therapeutic target of osteosarcoma. Cell Death & Disease, 13(11), 954.

Publication 2022

fluorescence microscopy F-7000

Assay Cellular Fluorescence Fluorescence dyes Fluorescence microscopy Genetic modifications Lipid peroxidation Thiobarbituric acid reactive substances

Corresponding Organization : Second Affiliated Hospital of Soochow University

Other organizations : Xuzhou Children Hospital, Jiangsu Province Hospital

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Variable analysis

independent variables

Applied genetic modifications to OS cells

dependent variables

Fluorescence intensity of DCF-DA or CellROX staining
Cellular lipid peroxidation levels measured by TBAR activity assay

control variables

Number of OS cells per well (0.5 × 10^5 cells per well)
Cultivation time periods (not explicitly mentioned)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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