FIG. 3.
ABIN-2 preferentially interacts with a p105/TPL-2 complex. (A and B) Duplicate cultures of 293 cells were cotransfected with vectors encoding ABIN2-FL and HA-p105 or Myc-TPL-2 or with EV. Cell lysates were prepared from each duplicate culture set using either buffer A (1% NP-40) or RIPA buffer, as indicated. Lysates were resolved by SDS-PAGE (10% acrylamide) and Western blotting (top blots). HA-p105 and Myc-TPL-2 mRNA levels in total RNA were assayed by semiquantitative RT-PCR (bottom blots). The 18S rRNA amplicon was used as an internal control. (C) 293 cells were cotransfected with vectors encoding HA-p105 and TPL-2 individually or together. Transfected proteins were affinity purified from cell lysates, prepared in 1% NP-40 buffer A, using GST-ABIN-21-429 fusion protein coupled to glutathione-Sepharose. Isolated proteins were resolved by SDS-PAGE (10% acrylamide) and Western blotting. α, anti.