Apoptosis Evaluation in Glioma Stem Cells

After CoCl₂ application or 14-3-3β siRNA transfection, GSC cell apoptosis rate was evaluated by annexin V-fluorescein isothiocyanate and propidium iodide staining (BD Biosciences) for 15 min at RT (25°C) in the dark and flow cytometry analysis (BD, FACSuite Research Assay Software). Bax/Bcl-2 assessment is a way of assessing cell apoptosis [46 (link), 47 (link)]. Thus, Bax/Bcl-2 protein expression and mRNA levels were also detected by western blotting or qPCR following 14-3-3β siRNA transfection combined with CoCl₂ treatment (anti-Bax and anti-Bcl-2, 1 : 1,000; Santa Cruz Biotechnology, Inc.). In addition, caspase-3 activity was measured to determine GSC cell apoptosis after above treatments by using a kit (Promega, Inc.) according to the manufacturer's instructions [48 (link)].

Free full text: Click here

Cao W., Zhou Q., Wang H., Rao W., Cheng G., Wang P., Guo S., Ren B, & Zhang J. (2022). Hypoxia Promotes Glioma Stem Cell Proliferation by Enhancing the 14-3-3β Expression via the PI3K Pathway. Journal of Immunology Research, 2022, 5799776.

Publication 2022

annexin V-fluorescein isothiocyanate and propidium iodide staining anti-Bax anti-Bcl-2

Annexin v Apoptosis Assay Bcl 2 Caspase 3 Cell Combined treatment Flow cytometry Fluorescein Isothiocyanate Mrna Promega Propidium iodide Sirna Transfection

Corresponding Organization : Chinese PLA General Hospital

Top 5 similar protocols

Variable analysis

independent variables

CoCl2 application
14-3-3β siRNA transfection

dependent variables

GSC cell apoptosis rate
Bax/Bcl-2 protein expression
Bax/Bcl-2 mRNA levels
Caspase-3 activity

control variables

Annexin V-FITC and propidium iodide staining for 15 min at RT (25°C) in the dark
Flow cytometry analysis

positive controls

Anti-Bax and anti-Bcl-2 antibodies (1:1,000; Santa Cruz Biotechnology, Inc.)

negative controls

Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!