Immunophenotypic Analysis of Stem/Progenitor Cells

After 7 days of the treatment with the MEISi-1 and MEISi-2, the UCB- and PB-MNCs were labeled with PE-conjugated anti-human CD34 (Biolegend, Cat.No. 343506), APC-conjugated anti-human CD133 (Miltenyibiotec, Order No. 130-090-826) antibodies as we have done previously^{14 (link),16 (link),25}. In addition, UCB mononuclear cells were treated with Aldefluor reagent according to the manufacturer’s manual (ALDEFLUOR™ Kit, Stemcell Technologies, Cat.No. 01700). The expressions of CD34, CD133 surface markers and ALDH (Aldehyde Dehydrogenase) enzyme activity in the labeled cells were analyzed by flow cytometry (BD FACSCalibur™, Cell Analyzer) as we have done previously^{14 (link),16 (link),25}.

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Turan R.D., Albayrak E., Uslu M., Siyah P., Alyazici L.Y., Kalkan B.M., Aslan G.S., Yucel D., Aksoz M., Tuysuz E.C., Meric N., Durdagi S., Gulbas Z, & Kocabas F. (2020). Development of Small Molecule MEIS Inhibitors that modulate HSC activity. Scientific Reports, 10, 7994.

Publication 2020

PE-conjugated anti-human CD34 APC-conjugated anti-human CD133 ALDEFLUOR™ Kit

After treatment Aldehyde dehydrogenase Antibodies Cell Enzyme activity Flow cytometry Human Stemcell

Corresponding Organization : Yeditepe University

Other organizations : Acıbadem University, Koç University, Max Planck Institute for Heart and Lung Research, University of Minnesota, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Bahçeşehir University

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Variable analysis

independent variables

Treatment with MEISi-1 and MEISi-2

dependent variables

Expression of CD34, CD133 surface markers
ALDH (Aldehyde Dehydrogenase) enzyme activity in the labeled cells

control variables

Labeling of UCB- and PB-MNCs with PE-conjugated anti-human CD34 antibody, APC-conjugated anti-human CD133 antibody
Treatment of UCB mononuclear cells with Aldefluor reagent

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

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