FIG 2.

SMCHD1 forms a functional homodimer. (A) Reciprocal immunoprecipitation and Western blotting of SMCHD1-HA and SMCHD1-FLAG from nuclear extracts of HEK293T cells cotransfected with SMCHD1 plasmids. In, input (10%); IP, immunoprecipitate (15%). (B) Silver-stained IP sample from TAP-tagged mouse SMCHD1 expressed in human HEK293T cells. Both human and mouse SMCHD1 peptides were detected at similar levels. (C) Anti-FLAG Western blotting of fractions following size exclusion chromatography of rSMCHD1. (D) Anti-FLAG Western blotting of fractions from sucrose gradient sedimentation of rSMCHD1. Molecular mass standards (kilodaltons) for panels C and D are labeled above each blot. (E) Electron microscopy images of negatively stained rSMCHD1. Selected images are further magnified and presented at the right. Bars, 100 nm (left image) and 10 nm (right images). (F) Schematic of the proposed homodimeric form.