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. 2020 Jul 23;9:e58571. doi: 10.7554/eLife.58571

Figure 5. Mcm2-WT does not rescue the Mcm2Δ127 replication defect.

(A) Standard DNA replication reaction using p1017 (4.8 kb) as template. Cdt1·MCM2-Δ127 and Cdt1·MCM2-WT were included at the MCM loading step at the indicated ratios; the total concentration of Cdt1·MCM was 80 nM in the Mcm2-7 loading reaction. (B) Quantification of total relative DNA synthesis in reactions of experiment in C. Bars represent the average of two independent experiments. (C) Lane traces of experiment in C.

Figure 5—source data 1. Figure 5A, autoradiograph.

Figure 5.

Figure 5—figure supplement 1. The effect of Cdt1·MCM and DDK concentrations on DNA replication in vitro.

Figure 5—figure supplement 1.

(A) Cdt1·MCM titration experiment using standard DNA replication conditions. Template: p1017 (4.8 kbp). Left: Reaction products were analyzed by denaturing agarose gel-electrophoresis and autoradiography. Right: Plot of total normalized DNA synthesis. (B) DDK titration experiment using standard DNA replication conditions, but Cdt1·MCM2-Δ127 in place of Cdt1·MCM2-WT. Template: p1017 (4.8 kbp). Left: Reaction products were analyzed by denaturing agarose gel-electrophoresis and autoradiography. Right: Plot of total normalized DNA synthesis.
Figure 5—figure supplement 1—source data 1. Figure 5—figure supplement 1A.
Figure 5—figure supplement 1—source data 2. Figure 5—figure supplement 1A.