Figure 2

Interaction of EMD and CXXC5. (a) To assess the intracellular localization of EMD and CXXC5 when co-synthesized, HEK293 cells were transiently transfected for 48 h with the expression vector bearing 3F-CXXC5 and HA-EMD cDNA. Cells were then subjected to ICC using the Flag (green channel) or the HA (red channel) antibody. DAPI was used for DNA staining. The scale bar is 10 μm. Inset indicates a section with higher magnification. (b,c) To examine the protein synthesis, HEK293 cells were transfected with the expression vector bearing (b) 3F-CXXC5 and/or HA-EMD cDNA; or (c) HA-CXXC5 and/or 3F-EMD cDNA for 48 h. The synthesis of proteins was assessed by WB using the HA or the Flag antibody. HDAC1 used as a loading control was probed with the HDAC1 antibody. Star denotes distinct EMD species (d,e). The nuclear extracts (500 μg) of transiently co-transfected HEK293 cells were subjected to Co-IP with the HA (d) or the isotype-matched IgG. 50 μg of nuclear lysates was used as input control. The precipitates were subjected to SDS-10%PAGE followed with WB using analyzed using the Flag (d) or the HA (e) antibody. Molecular masses (MM) in kDa are indicated.