Figure 2
From: EBNA3C interacts with Gadd34 and counteracts the unfolded protein response
EBNA3C interacts with Gadd34 by co-IP and co-purifies with Gadd34 by microcystin pulldown. a) IB4 cells (5 million for each treatment) were collected, lysed in isotonic 0.5% NP-40 buffer, and immune precipitations (IP) performed with either protein G alone (PG) or protein G with the addition of anti-EBNA3C (A10) sera (PG/anti-EBNA3C) Proteins were western blotted for Gadd34 (SC-H193), following IP. b) Burkitt's lymphoma BJAB cells or BJAB-EBNA3C cells (5 million for each treatment) were collected, lysed in isotonic 0.5% NP-40 buffer, containing additional 0.5% BSA, and incubated with the indicated sepharose beads conjugated to either protein G or microcystin LR. Lysates and beads were rotated for 1 hr at 4 degrees, extensively washed with PBS, and affinity purified proteins western blotted for the presence of EBNA3C (E3C) and Gadd34.