Figure 5: Defective deletional V(D)J recombination and abnormal CJ formation 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 pMX-DELCJ recombination substrate with components, intermediates and products as defined for pMX-INV in Fig. 4a. (b) v-abl pro-B cell lines containing pMX-DELCJ were treated for 72 h with ABLki with or without ATMki and assayed by Southern blotting; EcoRV digest—C4 probe. Experiments were repeated at least two times. (c) PCR analysis of pMX-DELCJ CJs 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. Experiments were repeated at least two times and PCR specificity was confirmed by Southern blotting (Supplementary Fig. 8). (d) Sanger sequencing of PCR amplified CJs from v-abl pro-B cells (see also Supplementary Table 2). Number of base-pair deletions is indicated on the y-axis. The horizontal bar represents the median. The total number of sequenced CJs (n) is indicated. Outliers (>25 bp deletion) are not shown but are included in calculation of the median. *0.01≤P<0.05, ***P<0.001.
