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. 2014 Oct 30;10(10):e1004474. doi: 10.1371/journal.ppat.1004474

Figure 8. Binding of purified p12 to N-MLV CA-coated lipid nanotubes.

Figure 8

Multimeric arrays of wild type (WT) and P1G mutant N-MLV CA were generated by immobilising the His-tagged purified proteins on lipid nanotubes comprising the Ni2+-chelating lipid, DGS-NTA. Purified wild type p12 (lanes 2–4) or p12 Mutant 6 (lanes 6–8) were incubated with lipid nanotubes coated with WT CA (lanes 2 and 6), P1G CA (lanes 3 and 7) or no tubes (lanes 4 and 8) prior to centrifugation through a sucrose cushion. The pelleted material was resuspended in SDS-PAGE sample buffer and probed for CA and p12 by immunoblotting, using appropriate antibodies. Input, lanes 1 and 5, represents 1/40th dilution of the purified p12 proteins before incubation with CA-coated lipid nanotubes.