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. 2014 May 13;289(27):19164–19179. doi: 10.1074/jbc.M114.561704

FIGURE 1.

FIGURE 1.

Co-immunoprecipitation of TDP-43 with Y2H interactors. A–E, HEK293E cells were co-transfected with mCherry-TDP-43 FL, CTF, or control vector (ctrl.) along with control vectors (Ø), FLAG-tagged UBPY (A), LSM6, MED6, UBE2E3 (B), RNF2 (C), RBM45 (D), or HA-tagged RACK1 (E). Then cells were lysed and subjected to immunoprecipitation with anti-FLAG (A–D) or anti-HA (E). Western blotting was performed with total cell lysates (Input) and immunoprecipitates (FLAG-IP and HA-IP, respectively). Proteins were detected with antibodies against TDP-43; DsRed (mCherry), FLAG, or HA; and UBPY as indicated. The endogenous TDP-43 bands confirm even input loading. F and G, immunoprecipitation with anti-FLAG beads from HEK293E cell lysates co-expressing FLAG-TDP-43 FL and either Myc-UBE2E3 (F) or Myc-UBPY (G). Total protein (Input) and immunoprecipitated proteins (FLAG-IP) were subjected to WB with antibodies against UBE2E3, UBPY, Myc, TDP-43, FLAG, and GAPDH as a loading control.