Protein Expression Analysis of Tumor Tissue

Total protein was extracted from the tumor tissue using RIPA protein extraction reagent containing phosphatase and protease inhibitors. The total protein concentration was measured using a BCA Protein Assay Kit. Then, the protein samples were mixed with a loading buffer (containing β-mercaptoethanol) and were heated at 100 ℃ for 10 min. Immunoblotting was performed using Bcl2-Associated X Protein (BAX) (Abcam, ab32503), B-cell lymphoma 2 (Bcl-2) (Abcam, 182858), SLC7A11 (Huabio, HA600097), GPX-4 (Bioss, 3884R), p53 (Proteintech, 60283-2-1g), GSK-3β (Huabio, ET1607-71), p-GSK-3β (Bioss, 2066R), Nrf-2 (Santa Cruz, 365949), cluster of differentiation 31 (CD31) (Huabio, ER31219), vascular endothelial growth factor-A (VEGFA) (Huabio, ET1604-28), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), Histone-H3 (Proteintech, 17168-1-AP), and β-actin (Proteintech, HRP-60008). Densitometry was performed using ImageJ software.

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Xu F., Zhang J., Ji L., Cui W., Cui J., Tang Z., Sun N., Zhang G., Guo M., Liu B, & Dong J. (2023). Inhibition of Non-small Cell Lung Cancer by Ferroptosis and Apoptosis Induction through P53 and GSK-3β/Nrf2 Signal Pathways using Qingrehuoxue Formula. Journal of Cancer, 14(3), 336-349.

Publication 2023

Bcl-2 SLC7A11 GPX-4 GSK-3β Histone-H3 β-actin

Actin Assay B cell lymphoma Buffer Densitometry Histone h3 Mercaptoethanol Mmp2 Mmp9 Nrf 2 Protease inhibitors Protein Protein phosphatase Ripa Tissue Tumor Vascular endothelial growth factor a

Corresponding Organization : Fudan University

Other organizations : Shandong University of Traditional Chinese Medicine, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Nanjing Drum Tower Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Total protein concentration
Protein expression levels of Bcl2-Associated X Protein (BAX), B-cell lymphoma 2 (Bcl-2), SLC7A11, GPX-4, p53, GSK-3β, p-GSK-3β, Nrf-2, cluster of differentiation 31 (CD31), vascular endothelial growth factor-A (VEGFA), MMP2, MMP9

control variables

Histone-H3
β-actin

positive controls

None explicitly mentioned

negative controls

None explicitly mentioned

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