Quantifying Cell Proliferation with EdU Staining

EdU staining was performed to visualize proliferating cells using a Click-iT^TM EdU Alexa Fluor® 488 Imaging kit (Invitrogen) according to the manufacturer's instructions. Cells were seeded into eight-well slides and cultured overnight. The following day, 10 µM EdU was added to the culture media and incubated for an hour. The cells were then fixed and permeabilized. Nuclei were counterstained with Hoechst 33342. Soft agar colony formation assays were performed as described in a previous study 16 (link).

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Song Y., Zhang L., Jiang Y., Hu T., Zhang D., Qiao Q., Wang R., Wang M, & Han S. (2019). MTBP regulates cell survival and therapeutic sensitivity in TP53 wildtype glioblastomas. Theranostics, 9(20), 6019-6030.

Publication 2019

Click-iTTM EdU Alexa Fluor® 488 Imaging kit

Agar Alexa fluor 488 Assays Cells Culture media Hoechst 33342 Nuclei

Corresponding Organization : First Hospital of China Medical University

Other organizations : Shanghai First People's Hospital, Shanghai Jiao Tong University

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Variable analysis

independent variables

Addition of 10 µM EdU to the culture media

dependent variables

Proliferating cells visualized through EdU staining

control variables

Cells seeded into eight-well slides and cultured overnight
Cells fixed and permeabilized
Nuclei counterstained with Hoechst 33342

controls

No positive or negative controls were explicitly mentioned in the provided information.

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