The FAM104 proteins VCF1/2 promote the nuclear localization of p97/VCP.

By contrast, a clear trend towards reduced growth was observed for the knockout cells in the presence of CB-5083.

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To further confirm these results, we used high-content microscopy of control and VCF1/2 double-knockout HeLa cells and found a statistically highly significant reduction of the ratio of nuclear to cytoplasmic p97 by 25% ( Figure 6—figure supplement 1D ), in excellent agreement with the confocal microscopy analyses.

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While this effect did not reach statistical significance at all CB-5083 concentrations, it did so at 25 nM, that is, at a concentration that had no effect in the absence of camptothecin pretreatment (see Figure 7B ).

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