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. 2016 Apr 4;17(5):671–681. doi: 10.15252/embr.201541455

Figure EV2. Analysis of the role of PALB2 phosphorylation in RAD51 foci formation.

Figure EV2

  1. Influence of PALB2 phosphorylation mutants on RAD51 foci formation following exposure to IR. U2OS cells were siRNA transfected to knock down endogenous PALB2, then transfected with FLAG‐HA WT, S59A, S157/376A, or TMA–PALB2. Twenty‐four hours following construct transfection cells were irradiated with 15 Gy of IR and fixed and processed for IF 2 h later. Transfected cells were identified by HA antibody reactivity and scored for RAD51 foci (WT n = 53, S59A n = 54, S157/376A n = 52, TMA n = 51) (P‐values: *P = 0.0257, **P = 0.0029, ***P = 0.0003, unpaired Student's t‐test). Transient expression of the PALB2 constructs was confirmed by Western blot probing for the HA tag. Scale bar = 5 μm.
  2. Proficiency of clonal PALB2 cell lines to form RAD51 foci in response to DNA damage. Cell lines were assessed for RAD51 foci formation without siRNA transfection and transgene induction. Cells were exposed to IR (15 Gy) or HU (2 mM) and processed for RAD51 foci as in Fig 3A and B, respectively. Representative images are shown. Scale bar = 5 μm.