Fig. 4

Trim28 interacts with Tas in vivo. a Scheme of Trim28 domains and truncated constructs used in this study. RBCC domain represent ring finger, B-box, zinc finger, coiled-coil domain at its amino-terminus; PB is a planthomeodomain and bromodomain at its carboxyl-terminus; and a domain for HP1 binding in the middle of entire gene. b Trim28 interacts with Tas in vivo. HT1080 cells were infected with PFV for 48 hpi. Forty-eight hours later, the infected HT1080 cells were lysed, and immunoprecipitation was performed with a negative control IgG or anti-Trim28 antibody. Then, the immunoprecipitates were detected by Western blot using anti-Trim28 and anti-Tas antibodies. HEK293T cells were co-transfected with Flag-Trim28 and HA-Tas. Forty-eight hours post transfection, the whole cell lysate was harvested and Tas was immunoprecipitated with anti-HA and normal IgG, which was used as negative control. Then, the immunoprecipitates were detected by western blotting using anti-HA and anti-Flag antibodies. c The RBCC domain of Trim28 interacts with Tas in vivo. The HEK293T cells were co-transfected with HA-Trim28-△RBCC/Trim28-△PB/Trim28-M and Flag-Tas, and 48 h post transfection, the whole cell lysate was harvested and immunoprecipitated with anti-HA and normal IgG, which was used as negative control. Then, the immunoprecipitates were detected by Western blot using anti-Flag, and the input sample were detected by Western blot using anti-HA antibody. d Tas was colocalized with Trim28 in the nucleus in vivo. pEGFP-N1-Tas was transfected into HT1080 or HEK293T cells, and 48 h after transfection, Trim28 was visualized with anti-Trim28, while the nuclei were visualized with DAPI staining. A merged image is shown on the bottom. Scale bar, 7.5 μm