NOT1 MIF4G is required for CAF1 interactions in vivo. (A) Schematic of the coimmunoprecipitation assay with HA-CAF1 as the bait protein. (B) Western blots with antibodies to proteins that interact with CAF1 by GST pull-down were conducted to test interaction by co-IP. CCR4 is a positive control, and eIF4E is a negative control. Western blots depict a single representative experiment, and three biological replicates were conducted. (C) Schematic of the coimmunoprecipitation assay with HA-MIF4G (the NOT1 MIF4G domain) as the bait protein. Both HA-MIF4G WT and a CAF1-binding deficient mutant of the MIF4G domain (HA-MIF4G mut) were used. myc-CAF1 was coexpressed as a control. (D) Western blots were conducted to test whether 4E-T and Xp54 interact with the NOT1 MIF4G domain. eIF4E was used as a negative control, and myc-CAF1 also served as a control. Western blots depict a single representative experiment, and three biological replicates were conducted.
