Extended Data Fig. 1: SDS–PAGE gel and SEC-MALS analysis of the E.DndFGH complex.
a, Gel filtration profile of His-DndF, His-DndG, His-DndH and the DndFGH complex of B7A. The E.DndFGH complex was prepared by mixing His-DndF (64 μM), His-DndG (64 μM) and His-DndH (32 μM). b, A gel image of the SDS–PAGE analysis of eluted protein fractions from the size-exclusion chromatography. Results are representative of three independent experiments. c, No obvious DNA was observed in the eluted protein fractions by agarose gel electrophoresis. To exclude the involvement of DNA in the formation of the DndFGH complex, the mixture of DndF, DndG and DndH was treated with DNase I (0.033 U/µl) at 37 °C for 10 min. DNase I is capable of digesting DNA up to 5 μg under our experimental conditions. Results are representative of three independent experiments. d, SEC-MALS analysis shows that the E.DndFGH complex elutes with a molecular mass of 434.03 kDa, consistent with a theoretical value of 433.44 kDa when DndF, DndG and DndH, as shown in (a), assemble into a complex in a molar ratio of 2:2:1. The left and right axes represent the molecular weight and the light scattering detector reading, respectively. The black curve represents the calculated molecular weight. Results are representative of three independent experiments.
