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. 2017 Nov 10;28(1):69–89. doi: 10.1038/cr.2017.139

Figure 9.

Figure 9

Trim25-Blm recruitment and ATR activation are less robust in differentiated cells than in ESCs during replication stress response. (A) NIH3T3 cells had lower levels of Blm and Trim25 on replication forks than ESCs. (B) Knockdown of Trim25 in NIH3T3 cells impaired the ubiquitination of Blm. This defect could be rescued by WT Trim25, but not ΔRING-Trim25. (C) Knockdown of Trim25 in NIH3T3 cells impaired the recruitment of Blm to replication forks. This defect was rescued by WT Trim25, but not ΔRING-Trim25. (D) Knockdown of Trim25 in NIH3T3 cells impaired stalled fork restart. This defect was rescued by WT Trim25, but not ΔRING-Trim25. (E) Co-expression of Filia and Floped in NIH3T3 cells stimulated the ubiquitination of Blm upon HU treatment. Knockdown of Trim25 suppressed this modification. (F) In NIH3T3 cells co-expressing Filia and Floped, the recruitment of Blm to replication forks was decreased by depletion of Trim25. (G) Co-expression of Filia and Floped in NIH3T3 cells significantly increased the potential of stalled fork restart in a Trim25-dependent manner. (H) Time-course comparison of ATR activities among ESCs, ESCs depleted of Filia and Floped (ESC-double KO), NIH3T3 cells, and NIH3T3 cells expressing both Filia and Floped (3T3-Filia-Floped). Data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.