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. 2016 Dec 20;2:16047. doi: 10.1038/celldisc.2016.47

Figure 6.

Figure 6

BLM complex and FANCM work in the same pathway to promote replication traverse of ICLs. (a) Patterns of replication tracts in the vicinity of Dig-TMP ICLs (red dot) on DNA fibers. The protocol of this assay is illustrated on the top. The sequence of the differentially colored CldU (purple) and IdU (green) tracks defines the direction of replication forks. (b) Frequency of patterns in wild-type DT40 cells, RMI2 knockout DT40 cells and RMI2 knockout cells complemented with either the wild-type RMI2 gene or the RMI2 with K121A point mutant. (n=61, 70, 64 in the wild-type cells; 51, 75, 69 in RMI2-deficient DT40 cells; 63, 54, 52 in RMI2-deficient cells complemented with either the wild-type; 46, 74, 60 in the RMI2 K121A point mutant cells. ‘n’ indicates the number of encounters analyzed in individual experiments) (c) Frequency of patterns in wild-type DT40 cells, BLM-deficient DT40 cells and BLM-deficient cells complemented with either the wild-type BLM gene or the BLM with K466A mutant. Three independent experiments were performed. (n=75, 83, 62 in the wild-type cells; 45, 63, 74 in BLM-deficient DT40 cells; 63, 64, 52 in BLM-deficient cells complemented with either the wild-type BLM; 66, 74, 51 in the BLM K466A point mutant cells). (d) Frequency of patterns in wild-type DT40 cells, BLM-deficient DT40 cells, FANCM-deficient cells expressing FANCM D203A mutant and BLM knockout DT40 cells expressing FANCM D203A mutant (n=61, 78, 84 in the wild-type cells; 81, 75, 68 in BLM-deficient DT40 cells; 73, 64, 62 in FANCM-D203A mutant cells; 66, 84, 70 in the BLM−/− /FANCM-D203A cells). Dig-TMP, digoxigenin-tagged trimethylpsoralen; ICL, interstrand crosslink; MMC, mitomycin C.