Next Article in Journal
High Sensitive Biosensors Based on the Coupling Between Surface Plasmon Polaritons on Titanium Nitride and a Planar Waveguide Mode
Next Article in Special Issue
Head-Mounted Display-Based Microscopic Imaging System with Customizable Field Size and Viewpoint
Previous Article in Journal
A Novel Smooth Variable Structure Smoother for Robust Estimation
Open AccessArticle

Optimization of Experimental Variables Influencing Apoptosome Biosensor in HEK293T Cells

1
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran P.O. Box 14115-175, Iran
2
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran P.O. Box 14115-175, Iran
*
Author to whom correspondence should be addressed.
Sensors 2020, 20(6), 1782; https://doi.org/10.3390/s20061782
Received: 23 January 2020 / Revised: 5 February 2020 / Accepted: 13 February 2020 / Published: 23 March 2020
(This article belongs to the Special Issue Micro/Nanosensors for Cellular/Tissue Measurement)
The apoptotic protease-activating factor 1 (Apaf-1) split luciferase biosensor has been used as a biological tool for the detection of early stage of apoptosis. The effect of doxorubicin in a cell-based assay and the addition of cytochrome c and ATP in a cell-free system have been used to test the functionality of the reporter for the detection of apoptosome formation. Here, our data established a drug- and cytochrome c/ATP-independent way of apoptosis induction relying on the expression of the biosensor itself to induce formation of apoptosome. Overexpression of Apaf-1 constructs led to increased split luciferase activity and caspase-3 activity in the absence of any drug treatment. Caspase-3 activity was significantly inhibited when caspase-9DN was co-overexpressed, while the activity of the Apaf1 biosensor was significantly increased. Our results show that the Apaf-1 biosensor does not detect etoposide-induced apoptosis. View Full-Text
Keywords: apoptosis; apoptosome; luciferase complementary assay apoptosis; apoptosome; luciferase complementary assay
Show Figures

Figure 1

MDPI and ACS Style

Oladzad, A.; Nikkhah, M.; Hosseinkhani, S. Optimization of Experimental Variables Influencing Apoptosome Biosensor in HEK293T Cells. Sensors 2020, 20, 1782.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop