Skip to main content
. 2008 Aug 13;17(2):205–212. doi: 10.1038/ejhg.2008.150

Figure 4.

Figure 4

(a) Visualization of V(D)J recombination positive cells expressing EGFP with fluorescence microscopy. EGFP-positive HEK 293 cells were detected 48 h after transient transfection with pWTRAG-1/2 expression constructs and reported plasmids (see arrow). When WTRAG-1 was replaced with the C2438T mutant RAG-1 construct, no EGFP-positive cells were observed. pE50HK transfected cells served as the EGFP and HA positive control. (b) V(D)J recombination efficiency was analyzed by scoring EGFP/HA-positive cells by FACS. Cotransfection of HEK 293 cells with pC2438T-RAG-1 vector and reporter system plasmids gives minimal V(D)J recombination (0.008%) relative to WTRAG-1 transfections. Experiments were repeated in triplicate. (c) Statistical analysis showed that there was a significant difference between the WTRAG-1 and the C2438T mutant RAG-1 with respect to V(D)J recombination efficiency (P<0.05). (d) C2438T mutant RAG-1 protein is expressed at significant levels as detected by western blot but at consistently lower levels than WTRAG-1.