Computational protocol: Antitumorigenic effect of interferon-β by inhibition of undifferentiated glioblastoma cells

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Protocol publication

[…] The neural stem cell marker CD133 was employed as a marker of GSCs. Furthermore, glial fibrillary acidic protein (GFAP) was used as a marker of astrocytes, and galactocerebroside C (GalC) was used as a marker of oligodendrocytes (,,–). We employed the following fluorescence conjugated monoclonal antibodies: anti-CD133 (CD133-PE, 130-080-801; Miltenyi Biotec, Auburn, CA, USA), anti-GFAP (anti-GFAP-Alexa Fluor 488, 561449; Becton-Dickinson, NJ, USA), and anti-GalC (antiGalC-Alexa Fluor 488, MAB342A4; Millipore, Temecula, CA, USA).The expressions of CD133, GFAP and GalC in 0222-GSC, S-BTC, Rev-GSC, Rev-GSC+IFN, and the 7 human malignant glioma cell lines were analyzed with a fluorescence-activated cell sorter (FACS). The fluorescence was measured using FACSCalibur flow cytometer (Becton-Dickinson, Franklin Lakes, NJ, USA), and the DNA histograms were analyzed with Flowjo software (BioLegend, San Diego, CA, USA).The FACS analyses were repeated at least 3 times in each experiment, and we confirmed that similar tendencies were obtained. [...] Statistical analyses were performed using the unpaired, Mann-Whitney U test. If the samples comprised more than three groups, the significance of the overall samples was evaluated by the Kruskal-Wallis test before evaluating the significant differences between pairs of groups by the Mann-Whitney U test. All quantitative data are presented as the means ± SE from at least six samples per data point. Statistical software IBM SPSS Statistics version 21.0 (International Business Machines Corp., Armonk, NY, USA) was employed for the data analysis. […]

Pipeline specifications

Software tools FlowJo, SPSS
Applications Miscellaneous, Flow cytometry
Organisms Homo sapiens
Diseases Glioblastoma, Glioma, Neoplasms