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. 2021 Feb 15;22(3):e52164. doi: 10.15252/embr.202052164

Figure 5. Mitotic CMG disassembly in mouse ES cells is dependent upon TRAIP E3 ligase activity but does not require association of TRAIP with PCNA.

Figure 5

  • A
    TRAIP co‐purifies with the CMG helicase from extracts of mouse ES cells. The asterisk indicates a non‐specific band.
  • B
    Mouse ES cells were processed as in Fig 3B. Arrows in (i) indicate accumulation of GFP‐SLD5 on heterochromatin during interphase, whereas arrows in (ii) denote GFP‐SLD5 on chromatin during mitotic metaphase. Scale bars correspond to 10 µm.
  • C
    Immunoblot illustrating the stable expression from the CAG promoter of the indicated forms of TRAIP at the ROSA26 locus in TRAIP−/− mouse ES cells. Asterisks indicate non‐specific bands.
  • D
    Quantification of the percentage of mitotic cells with GFP‐SLD5 on mitotic chromosomes, after inhibition of CUL2LRR1 as above, in cells with the indicated genotypes. Cells were fixed before analysis.

Data Information: In (D), data are presented as mean of three experiments ± SD.