Fig. 3

Physical interaction of RFC2 and RFC4 with Mus81-Mms4 or Mus81. a Purified recombinant Mus81-Mms4 containing His₆-tag on Mms4 was mixed with yeast cell extracts (150 μL) containing Flag-tagged RFC2 or RFC4 and captured on Ni-NTA (lanes 2, 4, 6, and 8). After 30 min, the supernatant was removed, the beads washed, and bound proteins eluted. The fraction containing unbound proteins (Unbound) and the SDS eluates (Bound) were analyzed on SDS-PAGE followed by immunoblotting using anti-Flag and anti-His antibodies. As a control, yeast cell extracts containing either Flag-tagged RFC2 or RFC4 were mixed with Ni-NTA beads in the absence of the Mus81 complex (lanes 1, 3, 5, and 7). The positions of individual proteins are marked on the side of the gel. b Interaction of RFC2 or RFC4 with Mus81 subunit. The assay was performed as shown in (a). c Interaction of RFC2 or RFC4 with Mus81 (1–319) fragment. The assay was performed as shown in (a)