Fig. 5. Negative effect of TRIP-Br1/XIAP on necroptosis and apoptosis.
(A) Cells (2 × 105) were cultured in complete media for 24 h after seeding 2 × 105 cells into a 35-mm confocal dish. Resulting cells were transiently transfected with siXIAP, using scRNA as a non-silencing control and incubated in media without serum (SS) for 24 h. Cells were observed under confocal microscope (magnification, ×200) after staining with Annexin V-EnzoGold (enhanced Cyanine-3) to detect apoptotic cells labeled yellow and necroptotic cells stained with 7-Aminoactinomycin D (7-AAD) were detectedin red as described in Materials and Methods section. (B) Results were semi-quantified using ImageJ. Data are presented as mean ± SD based on three independent experiments.