Cloning and characterization of SHD1. (A) Interaction of SHD1 and STATs by 2-hybrid assay. C-terminal truncated forms of various STATs were subcloned into pAS2-1 vector and examined for the interaction with SHD1. The interaction was verified by the β-gal assay. (B) Schematic structure of SHD1. Asterisks denote potential start codons. Dark gray areas indicate 2 LXXLL motifs. An area with oblique lines is SAC3-homology domain. (C,D) Expression of SHD1 in a variety of mouse tissues (C) and embryos (D). Sk muscle indicates skeletal muscle. (E) SHD1-L and SHD1-S proteins expressed in 293T cells. Cell lysates were resolved by SDS-PAGE and Western blot was performed with anti-HA antibody. (F) Intracellular localization of SHD1 by immunostaining. A vector expressing SHD1-HA or a mock-HA vector was transfected into Hela cells and immunostained with anti-HA antibody. Original magnification: ×600.