Figure 2

HBZ interacted with C/EBPα protein. (A) HBZ interacted with C/EBPα. 293T cells were cotransfected with mycHis-HBZ together with FLAG-C/EBPα. After 48 hours, cell lysates were subjected to immunoprecipitation using anti–c-Myc followed by immunoblotting using anti-FLAG. (B) HBZ co-localized with C/EBPα. Hela cells were transfected with mycHis-HBZ and FLAG-C/EBPα. HBZ was detected using anti–MYC Cy3 antibody (ii). C/EBPα was detected using anti–Flag-biotin and secondary Streptavidin-Alexa 488 antibody (i). The overlay of HBZ and C/EBPα is shown (iii). (C) HBZ decreased C/EBPα’s DNA binding capability. After transfection with mycHis-HBZ, FLAG-C/EBPα, and pC/EBPα-Luc for 48 hours, 293T cells were chromatin immunoprecipitated by anti-FLAG antibody. The precipitated DNAs and 1% of the input cell lysates were amplified by the pC/EBP-Luc specific primers. Expression of HBZ and C/EBPα was detected by Western blot (bottom panel). (D) HBZ could not repress the level of C/EBPα. 293T cells were transfected with expression vector of C/EBPα and various amounts of mycHis-HBZ. After 48 hours, the cell lysates were subjected to immunoblotting.