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

Figure EV3. Analysis of interactions between PALB2 versions and key HDR proteins, as well as immunoblots upon siRNA depletion of BLM and FBH1.

Figure EV3

  1. The PALB2 cell lines were untreated or treated with doxycycline followed by exposure to IR treatment (15 Gy, 2 h recovery). The exogenous FLAG/HA‐tagged PALB2 was immunoprecipitated with FLAG‐agarose and the IP and input samples were run on SDS–PAGE. The membranes were blotted with RAD51 and HA antibodies and vinculin was used as a loading control.
  2. The cells were treated and analyzed as in (A). Immunoblotting was performed with BRCA2, HA, and vinculin antibodies.
  3. 293T cells were transfected with empty vector (EV) or GFP‐BRCA1 together with FLAG‐/HA‐tagged WT, TMA, or TMD PALB2. Immunoprecipitation was performed using FLAG‐agarose, and the IP and input samples were probed with GFP and HA antibodies. Vinculin was used as a loading control.
  4. Representative immunoblot for Fig 3D showing depletion of FBH1 and BLM. The cell lysates were analyzed by Western blotting and probed with FBH1, BLM, and vinculin antibodies.