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. Author manuscript; available in PMC: 2019 Oct 4.
Published in final edited form as: Mol Cell. 2018 Sep 20;72(1):19–36.e8. doi: 10.1016/j.molcel.2018.08.027

Figure 5. SPOP cancer mutants disrupt phase separation and DAXX degradation.

Figure 5.

(A) SPOP cancer mutants are defective at phase separation in vitro. Fluorescence microscopy/DIC images of WT SPOP or cancer mutants as a function of H-cDAXX concentration. All samples contain 10% w/v ficoll 70, 500 nM ORG-SPOP construct and/or Rhodamine-cDAXX. Camera settings were optimized in samples containing ~1:1 molar ratios for each row.

(B) SPOP cancer mutants are defective at co-localization with DAXX in HeLa cells. SC-35 (magenta) was used as marker for nuclear speckles. Cells with SPOP-DAXX co-localization or lack thereof are indicated. (For co-staining with PML, see Fig. S5B.)

(C) SPOP cancer mutants co-localize with DAXX when expressed at high levels. Whisker plot showing the signal intensity of V5-SPOP or V5-F133V (red points) and GFP-DAXX (green points) from (B) in which the V5-SPOP construct and GFP-DAXX co-localize or fail to co-localize. Each point represents a single cell. Horizontal lines indicate the mean; error bars indicate SD.