EBNA2 and EBF1 form protein complexes in CBF1 proficient and deficient DG75 B cells.

<p>(A) DG75^doxHA-E2 CBF1 wt and CBF1 ko B cells or (B) DG75^doxHA-E2WW CBF1 wt B cells were transfected with EBF1 expression plasmids or empty vector controls. EBNA2 expression was induced by doxycycline (Dox) treatment directly after transfection or cells were left untreated. Total cellular extracts were harvested after 24 h and subjected to immunoprecipitation (IP) using an EBF specific antibody and then assayed by Western blot (WB) using EBF and EBNA2 specific antibodies. Total cell lysates (L) represent 1% of the cells used for IP (n = 2, one representative experiment is shown). (C) DG75 were transfected with EBF1 expression plasmids (+) or empty vector controls (-). Total cellular extracts were harvested 24 h post transfection, incubated with GST or GST-END domain fusion proteins coupled beads to pull down associated proteins. Western blot detection was performed with EBF and GST specific antibodies. GAPDH was used as an internal loading control (n = 2, one representative experiment is displayed).