Journal: Cell Death & Disease
Article Title: Improving the therapeutic potential of endostatin by fusing it with the BAX BH3 death domain
Figure Lengend Snippet: Internalization of ES, ES-BAX, ES-BAK, and ES-BAX-ES by endothelial cells. ( a ) C-PAE or NIH 3T3 cells were incubated at 37°C for 2 h with 10 μ g/ml of the biotynilated proteins: ES, ES-BAX, ES-BAK, or ES-BAX-ES. The cells were then fixed, permeabilized, and incubated with Alexa 488 conjugated with streptavidin. The nucleuses were stained with DAPI. The cells were analyzed under a fluorescence microscope. Scale bar: 10 μ m. ( b ) Western blot of endothelial C-PAE cells incubated with 10 μ g/ml of the indicated proteins at 37°C and detected by anti-ES antibody. Control (+): ES; control (−): lysate of cells incubated with ES that were immediately processed for western blot ( t =0 min)
Article Snippet: C-PAE or NIH 3T3 cells (ATCC, CCL-209 and CRL-1658, respectively; 5 × 104 ) were cultured in glass coverslips in EMEM medium (Life Technologies Corporation, Grand Island, NY, USA) supplemented with 2% FBS for 24 h. After 24 h of serum deprivation, the cells were incubated for 12 h in the same medium supplemented with 5 ng/ml bFGF.
Techniques: Incubation, Staining, Fluorescence, Microscopy, Western Blot