Extended Data Fig. 1: Assembly of the Rad24-RFC–9-1-1–DNA ternary complexes.
From: DNA is loaded through the 9-1-1 DNA checkpoint clamp in the opposite direction of the PCNA clamp
a) SDS-PAGE gels of purified Rad24-RFC and the 9-1-1 clamp. Similar gels were run >3 times. b) A raw micrograph of the mixture of Rad24-RFC with 9-1-1 and a 3′-recessed DNA substrate after incubation in an ice-water bath for 3 hr. 12,000 similar micrographs were recorded. c) 2D class averages. No ternary complex of Rad24-RFC–9-1-1–DNA was observed. Only the separate 9-1-1 ring and the Rad24-RFC particles were observed. d) A typical raw micrograph of the mixture of Rad24-RFC with 9-1-1 and a double-tailed DNA substrate containing both the 5′- and 3′- recessed DNA junctions. 14521 similar micrographs were recorded. e) Selected 2D class averages with different views, revealing the presence of the targeted Rad24-RFC–9-1-1 clamp–DNA ternary complexes. Scale bars are shown in the lower right corner in each panel.
