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. 2009 Jan 9;37(5):1452–1462. doi: 10.1093/nar/gkn1067

Figure 3.

Figure 3.

Southern blot analysis reveals the deletion of both JBP1 and JBP2. (A) DNA was isolated from WT bloodstream cells (WT427), and cells from which either one (JBP2−/+(H)) or both (JBP2−/− (H/H)) alleles of JBP2 were deleted as previously described (9). JBP1 was then deleted from this cell line by by LOH (JBP2−/− (H/H)/JBP1−/− (P/P)) and by conventional methods (JBP2−/− (H/H)/JBP1−/− (P/B)). DNA samples were digested with PstI and probed for the presence of the JBP2 (left) or JBP1 (right) open reading frame (ORF). A clear loss of a band correlating with JBP2 and JBP1 is apparent in cell lines from which all four JBP alleles were deleted, regardless of the method of generation. The 1 kb DNA size marker is indicated on the left. The WT gene fragments are indicated by the arrows on the right. A cross-reactive 3 kb band is visible on the JBP1 ORF hybridization. H, P and B refer to the hygomycin, puromycin and blasticidin resistance genes, respectively. (B) Map demonstrating area of JBP1 or JBP2 targeted by deletion constructs. Pst I, HindIII, Nar I restriction sites are indicated. Puro, puromycin resistance gene; Hygro, hygromycin resistance gene. The ORF probes used in panel A are indicated by the lines.