Figure 2: Impaired rearrangement at the endogenous Igk locus in Rag2c/c XLF−/− B cells.
From: RAG2 and XLF/Cernunnos interplay reveals a novel role for the RAG complex in DNA repair
(a) Schematic representation of the Igk locus. The relative positions of the EcoRI and SacI restriction sites and the JKIII probe are shown. Schematic representations of SacI/EcoRI digest fragments for normal Vk-Jk rearrangements (left) and the Jk1, Jk2, Jk4, and Jk5 coding ends (CE) (right) are represented. SacI/EcoRI digest fragments from Vk-Jk rearrangements are variable in size depending on the Vk and Jk gene segments used and the relative positions of EcoRI/SacI sites, whereas the size of Jk1, 2, 4 and 5 coding ends fragments are 1.8, 1.5, 0.8 and 0.5 kb, respectively. (b) Southern blot (using JKIII probe) of EcoRI/SacI digested DNA from v-abl pro-B cell lines treated for 72 h with ABLki (see also Supplementary Fig. 2). Arrows indicate normal rearrangements. Jk1–5 CEs are indicated in red. (c) Schematic representation of the Igk locus with position of primers (arrows) used to assay CJ and HJ formation during inversional IgkV6–23-J1 rearrangement. (d) Semi-quantitative nested PCR analysis of IgkV6–23-J1 CJ and HJ from indicated v-abl abl pro-B cell lines treated for 72 h with ABLki with or without ATMki. Il2 gene PCR was used as a loading control. The data presented are representative of at least two experiments using independent cell lines.
