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. 2015 Mar 12;5:9057. doi: 10.1038/srep09057

Figure 4.

Figure 4

(A & B). Immunoprecipitation analysis. (A): PtMCM immunoprecipitates were resolved by SDS-PAGE (10%) and analyzed by western blotting using anti-MCM (to detect immunoprecipitated MCM) or anti-GINS (to detect co-immunoprecipitating GINS) antibodies (1:1000 dilution). Full-length blots can be seen on-line in Supplementary Fig. S4A. (B): PtGINS immunoprecipitates were similarly analyzed. Full-length blots can be seen on-line in Supplementary Fig. S4B. Lanes 1: antibody control (bead-bound antibodies only and no lysate added); lanes 2: lysate incubated with beads only (with no antibody coupled to the beads); lanes 3: immunoprecipitation reaction; lanes 4: molecular weight marker; lanes 5: input lysate. (C). MCM pulldown experiment: Coomassie stain of SDS-PAGE analysis. Lane 1-input purified PtMCM; lane 2 – input purified PtGINS; lanes 3 to 8 – analysis of eluate fractions of pulldowns. Lanes 3 and 6 - recombinant MCM alone incubated with Strep -Tactin Superflow affinity beads; lanes 4 and 7- recombinant GINS alone incubated with Strep -Tactin Superflow affinity beads; lanes 5 and 8- recombinant MCM plus recombinant GINS incubated with Strep-Tactin Superflow affinity beads. Mr – molecular weight marker.