A-B) Immunoprecipitations were carried out either with an anti-Flag antibody on extracts of HEK-293 cells transduced with a Flag-mouse ZC3H4 expression vector (A) or with an anti-ZC3H4 rabbit polyclonal antibody on extracts from Raw264.7 mouse macrophages (B). Different parts of the western blot membrane were hybridized with the indicated antibodies. Data are representative of n=4 independent experiments. The position of molecular weight markers (kDa) is shown on the right. Uncropped images are available online as Source Data.
C) Upper panel: Schematic representation of (A) the full length human ZC3H4 protein and (B to E) its deletion mutants used in transfection and co-immunoprecipitation experiments. The ZC3H4 domains annotated in UniProt are shown. Bottom panel: lysates from HEK-293 cells, either untransfected (-) or transduced with the indicated Flag-ZC3H4 expression vectors (A-E) were used in co-immunoprecipitation experiments with an anti-Flag antibody. Inputs (left) and immunoprecipitates (right) were immunoblotted and probed with an anti-FLAG (top) or an anti-WDR82 (bottom) antibody as indicated. The position of molecular weight markers (kDa) is shown on the right. Uncropped images are available online as Source Data.
D) The Flag-tagged ZC3H4 C-terminal fragment (804-1303) was expressed in HeLa cells. Lysates were immunoprecipitated with an anti-ZC3H4 antibody directed against aa. 677-765 and blotted with anti-Flag or anti-WDR82 antibody. Inputs are shown on the left and molecular weight markers (kDa) on the right. Uncropped images are available online as Source Data.