Cancer Research Program, Rajiv Gandhi Centre for Biotechnology (Research Centre, University of Kerala), Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India.
Cancer Research Program, Rajiv Gandhi Centre for Biotechnology (Research Centre, University of Kerala), Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India.
Toxicol Lett. 2020 Jun 15;326:23-30. doi: 10.1016/j.toxlet.2020.02.011. Epub 2020 Feb 25.
Simultaneous detection of autophagy and apoptosis is important in drug discovery and signaling studies. Here we report, a real-time reporter cell line for the simultaneous detection of apoptosis and autophagy at single-cell level employing stable integration of two fluorescent protein reporters of apoptosis and autophagy. Cells stably expressing EGFP-LC3 fusion was developed initially as a marker for autophagy and subsequently stably expressed with inter-mitochondrial membrane protein SMAC with RFP fusion to detect mitochondrial permeabilization event of apoptosis. The cell lines faithfully reported the LC3 punctae formation and release of intermembrane proteins in response to diverse apoptotic and autophagic stimuli.
同时检测自噬和细胞凋亡在药物发现和信号研究中非常重要。在此，我们报告了一种实时报告细胞系，可在单细胞水平上同时检测细胞凋亡和自噬，该细胞系通过稳定整合细胞凋亡和自噬的两种荧光蛋白报告基因实现。最初，细胞中稳定表达 EGFP-LC3 融合蛋白作为自噬的标志物，随后，细胞中稳定表达含有红色荧光蛋白（RFP）的线粒体跨膜蛋白 SMAC 融合蛋白，以检测细胞凋亡中线粒体通透性改变的事件。该细胞系忠实地报告了 LC3 斑点的形成和细胞内膜蛋白的释放，以响应各种凋亡和自噬刺激。
