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As shown in Figure 5B, several forms of Maf1 were present in the starting material, and only the minor fastest migrating form was found in the anti-RPC1 immunoprecipitate.
At peak activity (Fig. 1 F; 38 min), cyclin B1-bound cdk1 was mostly in the fastest migrating form, dephosphorylated at inhibitory sites, while slower migrating cdk1 forms were present at earlier and later time points (Fig. 1. F; 40, 42, 44, 46).
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Treatment of lysates from mitotically arrested cells with λ phosphatase resulted in the complete loss of the slower migrating band and a concomitant increase in the faster migrating form, indicating that the observed retardation in electrophoretic mobility was indeed dependent on phosphorylation (Supplementary Figure 4C).
Although the intensity of both bands was reduced in the fogsk8 mutant, the faster migrating form appeared more severely affected (Figure 1E).
This is supported by our observations that, in the absence of myristoylation, Nkd1G2A interacts preferentially with a faster migrating form of Dvl and both remain uniformly distributed within the cytoplasm.
The monomer G-quadruplex appears in the gel as the fastest migrating band, then the nonfolded sequence, whereas dimers and tetramers are slower migrating forms.
Again, [CherryFP-F2A] migrated as a doublet, with the fastest migrating product corresponding to the CherryFP cleavage product - of uniform size across the gel tracks.
Taken with the complementation data, this result suggests that faster migrating, presumably activated, form of Osw3 is necessary for function and that Osw3 might activate itself by auto-cleavage of the pro-domain.
Using this approach, we found gradual accumulation of human PMP22 within affected mouse nerves, including a fast migrating incompletely processed ˜18 kDa form.
We found that phosphatase treatment returned the modified form of Exo1 to its faster migrating original form, indicating that Exo1 is indeed phosphorylated.
The presence of both slower and faster migrating complexes in EMSA assays of samples formed at 2 mM Mg2+, where AFM predicts mostly linear filaments, is an inconsistency in our results but could reflect formation of bridged filaments during electrophoresis in the altered environment of the gel.
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